Cyclic amines as bromodomain inhibitors

ABSTRACT

The present disclosure relates to compounds, which are useful for inhibition of BET protein function by binding to bromodomains, and their use in therapy.

This application is a divisional of U.S. patent application Ser. No.14/085,545, filed Nov. 20, 2013 (issued as U.S. Pat. No. 9,073,878 onJul. 7, 2015), which claims the benefit of U.S. Provisional ApplicationNo. 61/729,097, filed Nov. 21, 2012, all of which are incorporatedherein by reference in their entirety.

The present disclosure relates to novel compounds, pharmaceuticalcompositions containing such compounds, and their use in prevention andtreatment of diseases and conditions.

Post-translational modifications (PTMs) of histones are involved inregulation of gene expression and chromatin organization in eukaryoticcells. Histone acetylation at specific lysine residues is a PTM that isregulated by histone acetylases (HATs) and deacetylases (HDACs) [1].Small molecule inhibitors of HDACs and HATs are being investigated ascancer therapy [2-5]. Histone acetylation controls gene expression byrecruiting protein complexes that bind directly to acetylated lysine viabromodomains [6]. One such family, the bromodomain and extra terminaldomain (BET) proteins, comprises Brd2, Brd3, Brd4, and BrdT, each ofwhich contains two bromodomains in tandem that can independently bind toacetylated lysines, as reviewed in [7].

Interfering with BET protein interactions via bromodomain inhibitionresults in modulation of transcriptional programs that are oftenassociated with diseases characterized by dysregulation of cell cyclecontrol, inflammatory cytokine expression, viral transcription,hematopoietic differentiation, insulin transcription, and adipogenesis[8].

BET inhibitors are believed to be useful in the treatment of diseases orconditions related to systemic or tissue inflammation, inflammatoryresponses to infection or hypoxia, cellular activation andproliferation, lipid metabolism, fibrosis, and the prevention andtreatment of viral infections [8, 9].

Autoimmune diseases, which are often chronic and debilitating, are aresult of a dysregulated immune response, which leads the body to attackits own cells, tissues, and organs. Pro-inflammatory cytokinesincluding, IL-1β, TNF-α, IL-6, MCP-1, and IL-17 are overexpressed inautoimmune disease. IL-17 expression defines the T cell subset known asTh17 cells, which are differentiated, in part, by IL-6, and drive manyof the pathogenic consequences of autoimmune disease. Thus, theIL-6/Th17 axis represents an important, potentially druggable target inautoimmune disease therapy [10].

BET inhibitors are expected to have anti-inflammatory andimmunomodulatory properties [8, 9]. BET inhibitors have been shown tohave a broad spectrum of anti-inflammatory effects in vitro includingthe ability to decrease expression of pro-inflammatory cytokines such asIL-1β, MCP-1, TNF-α, and IL-6 in activated immune cells [11-13]. Themechanism for these anti-inflammatory effects may involve BET inhibitordisruption of Brd4 co-activation of NF-κB-regulated pro-inflammatorycytokines and/or displacement of BET proteins from cytokine promoters,including IL-6 [12, 14, 15]. In addition, because Brd4 is involved inT-cell lineage differentiation, BET inhibitors may be useful ininflammatory disorders characterized by specific programs of T celldifferentiation [16].

The anti-inflammatory and immunomodulatory effects of BET inhibitionhave also been confirmed in vivo. A BET inhibitor rescued mice fromendotoxin- or bacterial sepsis-induced death and cecal ligationpuncture-induced death, suggesting utility for BET inhibitors in sepsisand acute inflammatory disorders [12]. A BET inhibitor has been shown toameliorate inflammation and kidney injury in HIV-1 transgenic mice, ananimal model for HIV-associated nephropathy, in part through inhibitionof Brd4 interaction with NF-κB [14]. The utility of BET inhibition inautoimmune disease was demonstrated in a mouse model of multiplesclerosis, where BET inhibition resulted in abrogation of clinical signsof disease, in part, through inhibition of IL-6 and IL-17 [17]. Theseresults were supported in a similar mouse model where it was shown thattreatment with a BET inhibitor inhibited T cell differentiation intopro-autoimmune Th1 and Th17 subsets in vitro, and further abrogateddisease induction by pro-inflammatory Th1 cells [18].

BET inhibitors may be useful in the treatment of a variety of chronicautoimmune inflammatory conditions. Examples of autoimmune andinflammatory diseases, disorders, and syndromes treated using thecompounds and methods include, but are not limited to, inflammatorypelvic disease, urethritis, skin sunburn, sinusitis, pneumonitis,encephalitis, meningitis, myocarditis, nephritis [14], osteomyelitis,myositis, hepatitis, gastritis, enteritis, dermatitis, gingivitis,appendicitis, pancreatitis, cholecystitis, agammaglobulinemia,psoriasis, allergy, Crohn's disease, irritable bowel syndrome,ulcerative colitis [9], Sjogren's disease, tissue graft rejection,hyperacute rejection of transplanted organs, asthma, allergic rhinitis,chronic obstructive pulmonary disease (COPD), autoimmune polyglandulardisease (also known as autoimmune polyglandular syndrome), autoimmunealopecia, pernicious anemia, glomerulonephritis, dermatomyositis,multiple sclerosis [18], scleroderma, vasculitis, autoimmune hemolyticand thrombocytopenic states, Goodpasture's syndrome, atherosclerosis,Addison's disease, Parkinson's disease, Alzheimer's disease, Type Idiabetes [8], septic shock [12], systemic lupus erythematosus (SLE) [9],rheumatoid arthritis [19], psoriatic arthritis, juvenile arthritis,osteoarthritis, chronic idiopathic thrombocytopenic purpura, Waldenstrommacroglobulinemia, myasthenia gravis, Hashimoto's thyroiditis, atopicdermatitis, degenerative joint disease, vitiligo, autoimmunehypopituitarism, Guillain-Barre syndrome, Behcet's disease, uveitis, dryeye disease, scleroderma, mycosis fungoides, and Graves' disease.

BET inhibitors may be useful in the treatment of a wide variety of acuteinflammatory conditions including, but not limited to, acute gout, giantcell arteritis, nephritis including lupus nephritis, vasculitis withorgan involvement, such as glomerulonephritis, vasculitis, includinggiant cell arteritis, Wegener's granulomatosis, polyarteritis nodosa,Behcet's disease, Kawasaki disease, and Takayasu's arteritis.

BET inhibitors may be useful in the prevention and treatment of diseasesor conditions that involve inflammatory responses to infections withbacteria, viruses, fungi, parasites, and their toxins, such as, but notlimited to sepsis, sepsis syndrome, septic shock [12], systemicinflammatory response syndrome (SIRS), multi-organ dysfunction syndrome,toxic shock syndrome, acute lung injury, adult respiratory distresssyndrome (ARDS), acute renal failure, fulminant hepatitis, burns,post-surgical syndromes, sarcoidosis, Herxheimer reactions,encephalitis, myelitis, meningitis, malaria, and SIRS associated withviral infections, such as influenza, herpes zoster, herpes simplex, andcoronavirus [8].

Cancer is a group of diseases caused by dysregulated cell proliferation.Therapeutic approaches aim to decrease the numbers of cancer cells byinhibiting cell replication or by inducing cancer cell differentiationor death, but there is still significant unmet medical need for moreefficacious therapeutic agents. Cancer cells accumulate genetic andepigenetic changes that alter cell growth and metabolism, promoting cellproliferation and increasing resistance to programmed cell death, orapoptosis. Some of these changes include inactivation of tumorsuppressor genes, activation of oncogenes, and modifications of theregulation of chromatin structure, including deregulation of histonePTMs [20, 21].

The present disclosure provides a method for treating human cancer,including, but not limited to, cancers that result from aberranttranslocation or overexpression of BET proteins (e.g., NUT midlinecarcinoma (NMC) [22]) and B-cell lymphoma [23]). NMC tumor cell growthis driven by a translocation of the Brd4 or Brd3 gene to the nutlin 1gene [24]. BET inhibition has demonstrated potent antitumor activity inmurine xenograft models of NMC, a rare but lethal form of cancer [24].

The present disclosure provides a method for treating human cancers,including, but not limited to, cancers dependent on a member of the mycfamily of oncoproteins including c-myc, MYCN, and L-myc [25]. Thesecancers include Burkitt's lymphoma, acute myelogenous leukemia, multiplemyeloma, and aggressive human medulloblastoma [25]. Cancers in whichc-myc is overexpressed may be particularly susceptible to BET proteininhibition; it has been shown that treatment of tumors that haveactivation of c-myc with a BET inhibitor resulted in tumor regressionthrough inactivation of c-myc transcription [26-30].

The present disclosure provides a method for treating human cancersincluding cancers that rely on BET proteins and pTEFb (Cdk9/CyclinT) toregulate oncogenes [31], and cancers that can be treated by inducingapoptosis or senescence by inhibiting Bcl2, cyclin-dependent kinase 6(CDK6) [26], or human telomerase reverse transcriptase (hTERT) [27, 32].

BET inhibitors may be useful in the treatment of cancers including, butnot limited to, adrenal cancer, acinic cell carcinoma, acoustic neuroma,acral lentiginous melanoma, acrospiroma, acute eosinophilic leukemia,acute erythroid leukemia, acute lymphoblastic leukemia, acutemegakaryoblastic leukemia, acute monocytic leukemia, acute myeloidleukemia [26, 28, 30], adenocarcinoma, adenoid cystic carcinoma,adenoma, adenomatoid odontogenic tumor, adenosquamous carcinoma, adiposetissue neoplasm, adrenocortical carcinoma, adult T-cellleukemia/lymphoma, aggressive NK-cell leukemia, AIDS-related lymphoma,alveolar rhabdomyosarcoma, alveolar soft part sarcoma, ameloblasticfibroma, anaplastic large cell lymphoma, anaplastic thyroid cancer,angioimmunoblastic T-cell lymphoma, angiomyolipoma, angiosarcoma,astrocytoma, atypical teratoid rhabdoid tumor, B-cell acutelymphoblastic leukemia [29], B-cell chronic lymphocytic leukemia, B-cellprolymphocytic leukemia, B-cell lymphoma [23], basal cell carcinoma,biliary tract cancer, bladder cancer, blastoma, bone cancer, Brennertumor, Brown tumor, Burkitt's lymphoma [28], breast cancer, braincancer, carcinoma, carcinoma in situ, carcinosarcoma, cartilage tumor,cementoma, myeloid sarcoma, chondroma, chordoma, choriocarcinoma,choroid plexus papilloma, clear-cell sarcoma of the kidney,craniopharyngioma, cutaneous T-cell lymphoma, cervical cancer,colorectal cancer, Degos disease, desmoplastic small round cell tumor,diffuse large B-cell lymphoma, dysembryoplastic neuroepithelial tumor,dysgerminoma, embryonal carcinoma, endocrine gland neoplasm, endodermalsinus tumor, enteropathy-associated T-cell lymphoma, esophageal cancer,fetus in fetu, fibroma, fibrosarcoma, follicular lymphoma, follicular,thyroid cancer, ganglioneuroma, gastrointestinal cancer, germ celltumor, gestational choriocarcinoma, giant cell fibroblastoma, giant celltumor of the bone, glial tumor, glioblastoma multiforme, glioma,gliomatosis cerebri, glucagonoma, gonadoblastoma, granulosa cell tumor,gynandroblastoma, gallbladder cancer, gastric cancer, hairy cellleukemia, hemangioblastoma, head and neck cancer, hemangiopericytoma,hematological malignancy, hepatoblastoma, hepatosplenic T-cell lymphoma,Hodgkin's lymphoma, non-Hodgkin's lymphoma, invasive lobular carcinoma,intestinal cancer, kidney cancer, laryngeal cancer, lentigo maligna,lethal midline carcinoma, leukemia, leydig cell tumor, liposarcoma, lungcancer, lymphangioma, lymphangiosarcoma, lymphoepithelioma, lymphoma,acute lymphocytic leukemia, acute myelogenous leukemia [28], chroniclymphocytic leukemia, liver cancer, small cell lung cancer, non-smallcell lung cancer, MALT lymphoma, malignant fibrous histiocytoma,malignant peripheral nerve sheath tumor, malignant triton tumor, mantlecell lymphoma, marginal zone B-cell lymphoma, mast cell leukemia,mediastinal germ cell tumor, medullary carcinoma of the breast,medullary thyroid cancer, medulloblastoma, melanoma [33], meningioma,Merkel cell cancer, mesothelioma, metastatic urothelial carcinoma, mixedMullerian tumor, mixed lineage leukemia [26], mucinous tumor, multiplemyeloma [27], muscle tissue neoplasm, mycosis fungoides, myxoidliposarcoma, myxoma, myxosarcoma, nasopharyngeal carcinoma, neurinoma,neuroblastoma, neurofibroma, neuroma, nodular melanoma, NUT-midlinecarcinoma [24], ocular cancer, oligoastrocytoma, oligodendroglioma,oncocytoma, optic nerve sheath meningioma, optic nerve tumor, oralcancer, osteosarcoma, ovarian cancer, Pancoast tumor, papillary thyroidcancer, paraganglioma, pinealoblastoma, pineocytoma, pituicytoma,pituitary adenoma, pituitary tumor, plasmacytoma, polyembryoma,precursor T-lymphoblastic lymphoma, primary central nervous systemlymphoma, primary effusion lymphoma, primary peritoneal cancer, prostatecancer, pancreatic cancer, pharyngeal cancer, pseudomyxoma peritonei,renal cell carcinoma, renal medullary carcinoma, retinoblastoma,rhabdomyoma, rhabdomyosarcoma, Richter's transformation, rectal cancer,sarcoma, Schwannomatosis, seminoma, Sertoli cell tumor, sex cord-gonadalstromal tumor, signet ring cell carcinoma, skin cancer, small blue roundcell tumors, small cell carcinoma, soft tissue sarcoma, somatostatinoma,soot wart, spinal tumor, splenic marginal zone lymphoma, squamous cellcarcinoma, synovial sarcoma, Sezary's disease, small intestine cancer,squamous carcinoma, stomach cancer, testicular cancer, thecoma, thyroidcancer, transitional cell carcinoma, throat cancer, urachal cancer,urogenital cancer, urothelial carcinoma, uveal melanoma, uterine cancer,verrucous carcinoma, visual pathway glioma, vulvar cancer, vaginalcancer, Waldenstrom's macroglobulinemia, Warthin's tumor, and Wilms'tumor.

BET inhibitors may be useful in the treatment of benign proliferativeand fibrotic disorders, including benign soft tissue tumors, bonetumors, brain and spinal tumors, eyelid and orbital tumors, granuloma,lipoma, meningioma, multiple endocrine neoplasia, nasal polyps,pituitary tumors, prolactinoma, pseudotumor cerebri, seborrheickeratoses, stomach polyps, thyroid nodules, cystic neoplasms of thepancreas, hemangiomas, vocal cord nodules, polyps, and cysts, Castlemandisease, chronic pilonidal disease, dermatofibroma, pilar cyst, pyogenicgranuloma, juvenile polyposis syndrome, idiopathic pulmonary fibrosis,renal fibrosis, post-operative stricture, keloid formation, scleroderma,and cardiac fibrosis.

Cardiovascular disease (CVD) is the leading cause of mortality andmorbidity in the United States [34]. Atherosclerosis, an underlyingcause of CVD, is a multifactorial disease characterized by dyslipidemiaand inflammation. BET inhibitors are expected to be efficacious inatherosclerosis and associated conditions because of aforementionedanti-inflammatory effects as well as ability to increase transcriptionof ApoA-I, the major constituent of HDL [11, 35].

Up-regulation of ApoA-I is considered to be a useful strategy intreatment of atherosclerosis and CVD [36], BET inhibitors have beenshown to increase ApoA-I transcription and protein expression [11, 35].Resverlogix has also shown that BET inhibitors bind directly to BETproteins and inhibit their binding to acetylated histones at the ApoA-1promoter, suggesting the presence of a BET protein repression complex onthe ApoA-1 promoter, which can be functionally disrupted by BETinhibitors. It follows that, BET inhibitors may be useful in thetreatment of disorders of lipid metabolism via the regulation of ApoA-Iand HDL such as hypercholesterolemia, dyslipidemia, atherosclerosis[36], and Alzheimer's disease and other neurological disorders [37].

BET inhibitors may be useful in the prevention and treatment ofconditions associated with ischemia-reperfusion injury such as, but notlimited to, myocardial infarction, stroke, acute coronary syndromes [9],renal reperfusion injury, organ transplantation, coronary artery bypassgrafting, cardio-pulmonary bypass procedures, hypertension, pulmonary,renal, hepatic, gastro-intestinal, or peripheral limb embolism.

Obesity-associated inflammation is a hallmark of type II diabetes,insulin resistance, and other metabolic disorders [8, 19]. Consistentwith the ability of BET inhibitors to inhibit inflammation, genedisruption of Brd2 in mice ablates inflammation and protects animalsfrom obesity-induced insulin resistance [38]. It has been shown thatBrd2 interacts with PPARγ and opposes its transcriptional function.Knockdown of Brd2 in vitro promotes transcription of PPARγ-regulatednetworks, including those controlling adipogenesis [39]. In additionBrd2 is highly expressed in pancreatic β-cells and regulatesproliferation and insulin transcription [38]. Taken together, thecombined effects of BET inhibitors on inflammation and metabolismdecrease insulin resistance and may be useful in the treatment ofpre-diabetic and type II diabetic individuals as well as patients withother metabolic complications [8].

Host-encoded BET proteins have been shown to be important fortranscriptional activation and repression of viral promoters. Brd4interacts with the E2 protein of human papilloma virus (HPV) to enableE2 mediated transcription of E2-target genes [40]. Similarly, Brd2,Brd3, and Brd4 all bind to latent nuclear antigen 1 (LANA1), encoded byKaposi's sarcoma-associated herpes virus (KSHV), promotingLANA1-dependent proliferation of KSHV-infected cells [41]. A BETinhibitor has been shown to inhibit the Brd4-mediated recruitment of thetranscription elongation complex pTEFb to the Epstein-Barr virus (EBV)viral C promoter, suggesting therapeutic value for EBV-associatedmalignancies [42]. Also, a BET inhibitor reactivated HIV in models oflatent T cell infection and latent monocyte infection, potentiallyallowing for viral eradication by complementary anti-retroviral therapy[43-46].

BET inhibitors may be useful in the prevention and treatment ofepisome-based DNA viruses including, but not limited to, humanpapillomavirus, herpes virus, Epstein-Barr virus, human immunodeficiencyvirus [8], adenovirus, poxvirus, hepatitis B virus, and hepatitis Cvirus.

Some central nervous system (CNS) diseases are characterized bydisorders in epigenetic processes. Brd2 haplo-insufficiency has beenlinked to neuronal deficits and epilepsy [47]. SNPs in variousbromodomain-containing proteins have also been linked to mentaldisorders including schizophrenia and bipolar disorders [9]. Inaddition, the ability of BET inhibitors to increase ApoA-I transcriptionmay make BET inhibitors useful in Alzheimer's disease therapyconsidering the suggested relationship between increased ApoA-I andAlzheimer's disease and other neurological disorders [37].

BRDT is the testis-specific member of the BET protein family which isessential for chromatin remodeling during [48, 49]. Genetic depletion ofBRDT or inhibition of BRDT interaction with acetylated histones by a BETinhibitor resulted in a contraceptive effect in mice, which wasreversible when small molecule BET inhibitors were used [50, 51]. Thesedata suggest potential utility of BET inhibitors as a novel andefficacious approach to male contraception.

Monocyte chemotactic protein-1 (MCP-1, CCU) plays an important role incardiovascular disease [52]. MCP-1, by its chemotactic activity,regulates recruitment of monocytes from the arterial lumen to thesubendothelial space, where they develop into macrophage foam cells, andinitiate the formation of fatty streaks which can develop intoatherosclerotic plaque [53]. The critical role of MCP-1 (and its cognatereceptor CCR2) in the development of atherosclerosis has been examinedin various transgenic and knockout mouse models on a hyperlipidemicbackground [54-57]. These reports demonstrate that abrogation of MCP-1signaling results in decreased macrophage infiltration to the arterialwall and decreased atherosclerotic lesion development.

The association between MCP-1 and cardiovascular disease in humans iswell-established [52]. MCP-1 and its receptor are overexpressed byendothelial cells, smooth muscle cells, and infiltratingmonocytes/macrophages in human atherosclerotic plaque [58]. Moreover,elevated circulating levels of MCP-1 are positively correlated with mostcardiovascular risk factors, measures of coronary atherosclerosisburden, and the incidence of coronary heart disease (CHD) [59]. CHDpatients with among the highest levels of MCP-1 are those with acutecoronary syndrome (ACS) [60]. In addition to playing a role in theunderlying inflammation associated with CHD, MCP-1 has been shown to beinvolved in plaque rupture, ischemic/reperfusion injury, restenosis, andheart transplant rejection [52].

MCP-1 also promotes tissue inflammation associated with autoimmunediseases including rheumatoid arthritis (RA) and multiple sclerosis(MS). MCP-1 plays a role in the infiltration of macrophages andlymphocytes into the joint in RA, and is overexpressed in the synovialfluid of RA patients [61]. Blockade of MCP-1 and MCP-1 signaling inanimal models of RA have also shown the importance of MCP-1 tomacrophage accumulation and proinflammatory cytokine expressionassociated with RA [62-65].

Overexpression of MCP-1, in the brain, cerebrospinal fluid (CSF), andblood, has also been associated with chronic and acute MS in humans[66]. MCP-1 is overexpressed by a variety of cell types in the brainduring disease progression and contributes to the infiltration ofmacrophages and lymphocytes which mediate the tissue damage associatedwith MS [66]. Genetic depletion of MCP-1 or CCR2 in the experimentalautoimmune encephalomyelitis (EAE) mouse model, a model resembling humanMS, results in resistance to disease, primarily because of decreasedmacrophage infiltration to the CNS [67, 68].

Preclinical data have suggested that small- and large-moleculeinhibitors of MCP-1 and CCR2 have potential as therapeutic agents ininflammatory and autoimmune indications.

The present disclosure includes compounds that are useful for inhibitionof BET protein function by binding to bromodomains, and their use in thetreatment and prevention of diseases and conditions, including, but notlimited to, cancer, autoimmune, and cardiovascular diseases.

The first aspect of the present disclosure includes compounds of FormulaI and methods of administering a therapeutically effective amount ofthose compounds to a mammal (e.g., a human) in need thereof:

or a stereoisomer, tautomer, pharmaceutical acceptable salt, or hydratethereof,wherein:

W₁ is selected from N and CR₁;

W₂ is selected from N and CR₂;

W₃ is selected from N and CR₃;

W₄ is selected from N and CR₄;

each W may be the same or different from each other;

A is selected from N and CH;

R₁, R₂, R₃, and R₄ are each independently selected from hydrogen, alkyl,alkenyl, alkynyl, alkoxy, thioalkyl, aryloxy, aryl, amino, hydroxyl, andhalogen;

two adjacent substituents selected from R₁, R₂, R₃, and R₄ may beconnected in a 5- or 6-membered ring to form a bicyclic carbocycle orbicyclic heterocycle;

AR1 is a group selected from the following:

B is a group selected from the following:

each ring system may be substituted with one or more substituentsindependently selected from R₁₀ and R₁₁;

R₅ is selected from hydrogen, alkoxy, alkyl, thioalkyl, aryloxy, aryl,hydroxyl, and halogen;

R₆ is selected from hydrogen, alkoxy, alkyl, thioalkyl, aryloxy, aryl,and halogen;

R₇ is selected from hydrogen, alkyl, —SO2R₁₂, —C(O)NR₁₂R₁₃, —C(O)R₁₂;

R₈ and R₉ are independently selected from hydrogen, aryl, alkenyl,alkyl, —SO₂R₁₂, —C(O)NR₁₂R₁₃, —C(O)R₁₂;

R₁₀ and R₁₁ are independently selected from hydrogen, halogen, alkyl,alkoxy, aryl, and hydroxyl;

R₁₂ and R₁₃ are independently selected from hydrogen, aryl, and alkyl;

Y is selected from NH, O, and S; and

two adjacent substituents selected from R₅, R₆, R₈, R₉, R₁₀, and R₁₁ maybe connected in a 5- or 6-membered ring to form a carbocycle orheterocycle.

In another aspect of the present disclosure, there is provided apharmaceutical composition comprising a compound of Formula I, or apharmaceutically acceptable salt thereof and one or morepharmaceutically acceptable carriers, diluents or excipients.

In yet another aspect of the present disclosure there is provided acompound of Formula I, or a pharmaceutically acceptable salt thereof foruse in therapy, in particular in the treatment of diseases or conditionsfor which a bromodomain inhibitor is indicated.

In yet another aspect of the present disclosure there is provided acompound of Formula I, or a pharmaceutically acceptable salt thereof inthe manufacture of a medicament for the treatment of diseases orconditions for which a bromodomain inhibitor is indicated.

DEFINITIONS

As used in the present specification, the following words, phrases andsymbols are generally intended to have the meanings as set forth below,except to the extent that the context in which they are used indicatesotherwise. The following abbreviations and terms have the indicatedmeanings throughout:

As used herein, “cardiovascular disease” refers to diseases, disordersand conditions of the heart and circulatory system that are mediated byBET inhibition. Exemplary cardiovascular diseases, includingcholesterol- or lipid-related disorders, include, but are not limitedto, acute coronary syndrome, angina, arteriosclerosis, atherosclerosis,carotid atherosclerosis, cerebrovascular disease, cerebral infarction,congestive heart failure, congenital heart disease, coronary heartdisease, coronary artery disease, coronary plaque stabilization,dyslipidemias, dyslipoproteinemias, endothelium dysfunctions, familialhypercholesterolemia, familial combined hyperlipidemia,hypoalphalipoproteinemia, hypertriglyceridemia,hyperbetalipoproteinemia, hypercholesterolemia, hypertension,hyperlipidemia, intermittent claudication, ischemia, ischemiareperfusion injury, ischemic heart diseases, cardiac ischemia, metabolicsyndrome, multi-infarct dementia, myocardial infarction, obesity,peripheral vascular disease, reperfusion injury, restenosis, renalartery atherosclerosis, rheumatic heart disease, stroke, thromboticdisorder, transitory ischemic attacks, and lipoprotein abnormalitiesassociated with Alzheimer's disease, obesity, diabetes mellitus,syndrome X, impotence, multiple sclerosis, Parkinson's disease, andinflammatory diseases.

As used herein, “inflammatory diseases” refers to diseases, disorders,and conditions that are mediated by BET inhibition. Exemplaryinflammatory diseases, include, but are not limited to, arthritis,asthma, dermatitis, psoriasis, cystic fibrosis, post transplantationlate and chronic solid organ rejection, multiple sclerosis, systemiclupus erythematosus, inflammatory bowel diseases, autoimmune diabetes,diabetic retinopathy, diabetic nephropathy, diabetic vasculopathy,ocular inflammation, uveitis, rhinitis, ischemia-reperfusion injury,post-angioplasty restenosis, chronic obstructive pulmonary disease(COPD), glomerulonephritis, Graves disease, gastrointestinal allergies,conjunctivitis, atherosclerosis, coronary artery disease, angina, andsmall artery disease.

As used herein, “cancer” refers to diseases, disorders, and conditionsthat are mediated by BET inhibition. Exemplary cancers, include, but arenot limited to, chronic lymphocytic leukemia and multiple myeloma,follicular lymphoma, diffuse large B cell lymphoma with germinal centerphenotype, Burkitt's lymphoma, Hodgkin's lymphoma, follicular lymphomasand activated, anaplastic large cell lymphoma, neuroblastoma and primaryneuroectodermal tumor, rhabdomyosarcoma, prostate cancer, breast cancer,NMC (NUT-midline carcinoma), acute myeloid leukemia (AML), acute Blymphoblastic leukemia (B-ALL), Burkitt's Lymphoma, B-cell lymphoma,melanoma, mixed lineage leukemia, multiple myeloma, pro-myelocyticleukemia (PML), non-Hodgkin's lymphoma, neuroblastoma, medulloblastoma,lung carcinoma (NSCLC, SCLC), and colon carcinoma.

“Subject” refers to an animal, such as a mammal, that has been or willbe the object of treatment, observation, or experiment. The methodsdescribed herein may be useful for both human therapy and veterinaryapplications. In one embodiment, the subject is a human.

As used herein, “treatment” or “treating” refers to an amelioration of adisease or disorder, or at least one discernible symptom thereof. Inanother embodiment, “treatment” or “treating” refers to an ameliorationof at least one measurable physical parameter, not necessarilydiscernible by the patient. In yet another embodiment, “treatment” or“treating” refers to inhibiting the progression of a disease ordisorder, either physically, e.g., stabilization of a discerniblesymptom, physiologically, e.g., stabilization of a physical parameter,or both. In yet another embodiment, “treatment” or “treating” refers todelaying the onset of a disease or disorder. For example, treating acholesterol disorder may comprise decreasing blood cholesterol levels.

As used herein, “prevention” or “preventing” refers to a reduction ofthe risk of acquiring a given disease or disorder.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example, —CONH₂ isattached through the carbon atom.

By “optional” or “optionally” is meant that the subsequently describedevent or circumstance may or may not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which is does not. For example, “optionally substituted aryl”encompasses both “aryl” and “substituted aryl” as defined below. It willbe understood by those skilled in the art, with respect to any groupcontaining one or more substituents, that such groups are not intendedto introduce any substitution or substitution patterns that aresterically impractical, synthetically non-feasible and/or inherentlyunstable.

As used herein, the term “hydrate” refers to a crystal form with eithera stoichiometric or non-stoichiometric amount of water is incorporatedinto the crystal structure.

The term “alkenyl” as used herein refers to an unsaturated straight orbranched hydrocarbon having at least one carbon-carbon double bond, suchas a straight or branched group of 2-8 carbon atoms, referred to hereinas (C₂-C₈)alkenyl. Exemplary alkenyl groups include, but are not limitedto, vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl,hexadienyl, 2-ethylhexenyl, 2-propyl-2-butenyl, and4-(2-methyl-3-butene)-pentenyl.

The term “alkoxy” as used herein refers to an alkyl group attached to anoxygen (—O-alkyl-). “Alkoxy” groups also include an alkenyl groupattached to an oxygen (“alkenyloxy”) or an alkynyl group attached to anoxygen (“alkynyloxy”) groups. Exemplary alkoxy groups include, but arenot limited to, groups with an alkyl, alkenyl or alkynyl group of 1-8carbon atoms, referred to herein as (C₁-C₈)alkoxy. Exemplary alkoxygroups include, but are not limited to methoxy and ethoxy.

The term “alkyl” as used herein refers to a saturated straight orbranched hydrocarbon, such as a straight or branched group of 1-8 carbonatoms, referred to herein as (C₁-C₈)alkyl. Exemplary alkyl groupsinclude, but are not limited to, methyl, ethyl, propyl, isopropyl,2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl,3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl,2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl,2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl,2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl,isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, andoctyl.

The term “alkynyl” as used herein refers to an unsaturated straight orbranched hydrocarbon having at least one carbon-carbon triple bond, suchas a straight or branched group of 2-8 carbon atoms, referred to hereinas (C₂-C₈)alkynyl. Exemplary alkynyl groups include, but are not limitedto, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl,4-methyl-1-butynyl, 4-propyl-2-pentynyl, and 4-butyl-2-hexynyl.

The term “amide” as used herein refers to the form —NR_(a)C(O)(R_(b))—or —C(O)NR_(b)R_(c), wherein R_(a), R_(b) and R_(c) are eachindependently selected from alkyl, alkenyl, alkynyl, aryl, arylalkyl,cycloalkyl, haloalkyl, heteroaryl, heterocyclyl, and hydrogen. The amidecan be attached to another group through the carbon, the nitrogen,R_(b), or R_(c). The amide also may be cyclic, for example R_(b) andR_(c), may be joined to form a 3- to 8-membered ring, such as 5- or6-membered ring. The term “amide” encompasses groups such assulfonamide, urea, ureido, carbamate, carbamic acid, and cyclic versionsthereof. The term “amide” also encompasses an amide group attached to acarboxy group, e.g., -amide-COOH or salts such as -amide-COONa, an aminogroup attached to a carboxy group (e.g., -amino-COOH or salts such as-amino-COONa).

The term “amine” or “amino” as used herein refers to the form—NR_(d)R_(e) or —N(R_(d))R_(e)—, where R_(d) and R_(e) are independentlyselected from alkyl, alkenyl, alkynyl, aryl, arylalkyl, carbamate,cycloalkyl, haloalkyl, heteroaryl, heterocyclyl, and hydrogen. The aminocan be attached to the parent molecular group through the nitrogen. Theamino also may be cyclic, for example any two of R_(d) and R_(e) may bejoined together or with the N to form a 3- to 12-membered ring (e.g.,morpholino or piperidinyl). The term amino also includes thecorresponding quaternary ammonium salt of any amino group. Exemplaryamino groups include alkylamino groups, wherein at least one of R_(d) orR_(e) is an alkyl group. In some embodiments Rd and Re each may beoptionally substituted with hydroxyl, halogen, alkoxy, ester, or amino.

The term “aryl” as used herein refers to a mono-, bi-, or othermulti-carbocyclic, aromatic ring system. The aryl group can optionallybe fused to one or more rings selected from aryls, cycloalkyls, andheterocyclyls. The aryl groups of this present disclosure can besubstituted with groups selected from alkoxy, aryloxy, alkyl, alkenyl,alkynyl, amide, amino, aryl, arylalkyl, carbamate, carboxy, cyano,cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl,heterocyclyl, hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl,sulfonyl, sulfonic acid, sulfonamide, and thioketone. Exemplary arylgroups include, but are not limited to, phenyl, tolyl, anthracenyl,fluorenyl, indenyl, azulenyl, and naphthyl, as well as benzo-fusedcarbocyclic moieties such as 5,6,7,8-tetrahydronaphthyl. Exemplary arylgroups also include, but are not limited to a monocyclic aromatic ringsystem, wherein the ring comprises 6 carbon atoms, referred to herein as“(C₆)aryl.”

The term “arylalkyl” as used herein refers to an alkyl group having atleast one aryl substituent (e.g., -aryl-alkyl-). Exemplary arylalkylgroups include, but are not limited to, arylalkyls having a monocyclicaromatic ring system, wherein the ring comprises 6 carbon atoms,referred to herein as “(C₆)arylalkyl.”

The term “carbamate” as used herein refers to the form—R_(g)OC(O)N(R_(h))—, —R_(g)OC(O)N(R_(h))R_(i)—, or —OC(O)NR_(h)R_(i),wherein R_(g), R_(h) and R_(i) are each independently selected fromalkyl, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, haloalkyl,heteroaryl, heterocyclyl, and hydrogen. Exemplary carbamates include,but are not limited to, arylcarbamates or heteroaryl carbamates (e.g.,wherein at least one of R_(g), R_(h) and R_(i) are independentlyselected from aryl or heteroaryl, such as pyridine, pyridazine,pyrimidine, and pyrazine).

The term “carboxy” as used herein refers to —COOH or its correspondingcarboxylate salts (e.g., —COONa). The term carboxy also includes“carboxycarbonyl,” e.g. a carboxy group attached to a carbonyl group,e.g., —C(O)—COOH or salts, such as —C(O)—COONa.

The term “cyano” as used herein refers to —CN.

The term “cycloalkoxy” as used herein refers to a cycloalkyl groupattached to an oxygen.

The term “cycloalkyl” as used herein refers to a saturated orunsaturated cyclic, bicyclic, or bridged bicyclic hydrocarbon group of3-12 carbons, or 3-8 carbons, referred to herein as “(C₃-C₈)cycloalkyl,”derived from a cycloalkane. Exemplary cycloalkyl groups include, but arenot limited to, cyclohexanes, cyclohexenes, cyclopentanes, andcyclopentenes. Cycloalkyl groups may be substituted with alkoxy,aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl,carbamate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen,haloalkyl, heteroaryl, heterocyclyl, hydroxyl, ketone, nitro, phosphate,sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide and thioketone.Cycloalkyl groups can be fused to other cycloalkyl saturated orunsaturated, aryl, or heterocyclyl groups.

The term “dicarboxylic acid” as used herein refers to a group containingat least two carboxylic acid groups such as saturated and unsaturatedhydrocarbon dicarboxylic acids and salts thereof. Exemplary dicarboxylicacids include alkyl dicarboxylic acids. Dicarboxylic acids may besubstituted with alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino,aryl, arylalkyl, carbamate, carboxy, cyano, cycloalkyl, ester, ether,formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydrogen,hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl, sulfonyl,sulfonic acid, sulfonamide and thioketone. Dicarboxylic acids include,but are not limited to succinic acid, glutaric acid, adipic acid,suberic acid, sebacic acid, azelaic acid, maleic acid, phthalic acid,aspartic acid, glutamic acid, malonic acid, fumaric acid, (+)/(−)-malicacid, (+)/(−) tartaric acid, isophthalic acid, and terephthalic acid.Dicarboxylic acids further include carboxylic acid derivatives thereof,such as anhydrides, imides, hydrazides (for example, succinic anhydrideand succinimide).

The term “ester” refers to the structure —C(O)O—, —C(O)O—R_(j)—,—R_(k)C(O)O—R_(j)—, or —R_(k)C(O)O—, where O is not bound to hydrogen,and R_(j) and R_(k) can independently be selected from alkoxy, aryloxy,alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, cycloalkyl,ether, haloalkyl, heteroaryl, and heterocyclyl. R_(k) can be a hydrogen,but R_(j) cannot be hydrogen. The ester may be cyclic, for example thecarbon atom and R_(j), the oxygen atom and R_(k), or R_(j) and R_(k) maybe joined to form a 3- to 12-membered ring. Exemplary esters include,but are not limited to, alkyl esters wherein at least one of Rj or Rk isalkyl, such as —O—C(O)-alkyl, —C(O)—O-alkyl-, and -alkyl-C(O)—O-alkyl-.Exemplary esters also include aryl or heteroaryl esters, e.g. wherein atleast one of Rj or Rk is a heteroaryl group such as pyridine,pyridazine, pyrimidine and pyrazine, such as a nicotinate ester.Exemplary esters also include reverse esters having the structure—R_(k)C(O)O—, where the oxygen is bound to the parent molecule.Exemplary reverse esters include succinate, D-argininate, L-argininate,L-lysinate and D-lysinate. Esters also include carboxylic acidanhydrides and acid halides.

The terms “halo” or “halogen” or “Hal” as used herein refer to F, Cl,Br, or I.

The term “haloalkyl” as used herein refers to an alkyl group substitutedwith one or more halogen atoms. “Haloalkyls” also encompass alkenyl oralkynyl groups substituted with one or more halogen atoms.

The term “heteroaryl” as used herein refers to a mono-, bi-, ormulti-cyclic, aromatic ring system containing one or more heteroatoms,for example 1-3 heteroatoms, such as nitrogen, oxygen, and sulfur.Heteroaryls can be substituted with one or more substituents includingalkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl,carbamate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen,haloalkyl, heteroaryl, heterocyclyl, hydroxyl, ketone, nitro, phosphate,sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide and thioketone.Heteroaryls can also be fused to non-aromatic rings. Illustrativeexamples of heteroaryl groups include, but are not limited to,pyridinyl, pyridazinyl, pyrimidyl, pyrazyl, triazinyl, pyrrolyl,pyrazolyl, imidazolyl, (1,2,3)- and (1,2,4)-triazolyl, pyrazinyl,pyrimidilyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, furyl,phenyl, isoxazolyl, and oxazolyl. Exemplary heteroaryl groups include,but are not limited to, a monocyclic aromatic ring, wherein the ringcomprises 2-5 carbon atoms and 1-3 heteroatoms, referred to herein as“(C₂-C₅)heteroaryl.”

The terms “heterocycle,” “heterocyclyl,” or “heterocyclic” as usedherein refer to a saturated or unsaturated 3-, 4-, 5-, 6- or 7-memberedring containing one, two, or three heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur. Heterocycles can be aromatic(heteroaryls) or non-aromatic. Heterocycles can be substituted with oneor more substituents including alkoxy, aryloxy, alkyl, alkenyl, alkynyl,amide, amino, aryl, arylalkyl, carbamate, carboxy, cyano, cycloalkyl,ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl,hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl, sulfonyl,sulfonic acid, sulfonamide and thioketone. Heterocycles also includebicyclic, tricyclic, and tetracyclic groups in which any of the aboveheterocyclic rings is fused to one or two rings independently selectedfrom aryls, cycloalkyls, and heterocycles. Exemplary heterocyclesinclude acridinyl, benzimidazolyl, benzofuryl, benzothiazolyl,benzothienyl, benzoxazolyl, biotinyl, cinnolinyl, dihydrofuryl,dihydroindolyl, dihydropyranyl, dihydrothienyl, dithiazolyl, furyl,homopiperidinyl, imidazolidinyl, imidazolinyl, imidazolyl, indolyl,isoquinolyl, isothiazolidinyl, isothiazolyl, isoxazolidinyl, isoxazolyl,morpholinyl, oxadiazolyl, oxazolidinyl, oxazolyl, piperazinyl,piperidinyl, pyranyl, pyrazolidinyl, pyrazinyl, pyrazolyl, pyrazolinyl,pyridazinyl, pyridyl, pyrimidinyl, pyrimidyl, pyrrolidinyl,pyrrolidin-2-onyl, pyrrolinyl, pyrrolyl, quinolinyl, quinoxaloyl,tetrahydrofuryl, tetrahydroisoquinolyl, tetrahydropyranyl,tetrahydroquinolyl, tetrazolyl, thiadiazolyl, thiazolidinyl, thiazolyl,thienyl, thiomorpholinyl, thiopyranyl, and triazolyl.

The terms “hydroxy” and “hydroxyl” as used herein refers to —OH.

The term “hydroxyalkyl” as used herein refers to a hydroxy attached toan alkyl group.

The term “hydroxyaryl” as used herein refers to a hydroxy attached to anaryl group.

The term “ketone” as used herein refers to the structure —C(O)—Rn (suchas acetyl, —C(O)CH₃) or —R_(n)—C(O)—R_(o)—. The ketone can be attachedto another group through Rn or R_(o). Rn or R_(o) can be alkyl, alkenyl,alkynyl, cycloalkyl, heterocyclyl or aryl, or Rn or R_(o) can be joinedto form a 3- to 12-membered ring.

The term “monoester” as used herein refers to an analogue of adicarboxylic acid wherein one of the carboxylic acids is functionalizedas an ester and the other carboxylic acid is a free carboxylic acid orsalt of a carboxylic acid. Examples of monoesters include, but are notlimited to, to monoesters of succinic acid, glutaric acid, adipic acid,suberic acid, sebacic acid, azelaic acid, oxalic and maleic acid.

The term “phenyl” as used herein refers to a 6-membered carbocyclicaromatic ring. The phenyl group can also be fused to a cyclohexane orcyclopentane ring. Phenyl can be substituted with one or moresubstituents including alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide,amino, aryl, arylalkyl, carbamate, carboxy, cyano, cycloalkyl, ester,ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl,ketone, nitro, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid,sulfonamide and thioketone.

The term “thioalkyl” as used herein refers to an alkyl group attached toa sulfur (—S-alkyl-).

“Alkyl,” “alkenyl,” “alkynyl”, “alkoxy”, “amino” and “amide” groups canbe optionally substituted with or interrupted by or branched with atleast one group selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl,amide, amino, aryl, arylalkyl, carbamate, carbonyl, carboxy, cyano,cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl,heterocyclyl, hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl,sulfonyl, sulfonic acid, sulfonamide, thioketone, ureido and N. Thesubstituents may be branched to form a substituted or unsubstitutedheterocycle or cycloalkyl.

As used herein, a “suitable substituent” refers to a group that does notnullify the synthetic or pharmaceutical utility of the compounds of thepresent disclosure or the intermediates useful for preparing them.Examples of suitable substituents include, but are not limited to: C₁₋₈alkyl, alkenyl or alkynyl; C₁₋₆ aryl, C₂₋₅ heteroaryl; C₃₋₇ cycloalkyl;C₁₋₈ alkoxy; C₆ aryloxy; —CN; —OH; oxo; halo, carboxy; amino, such as—NH(C₁₋₈ alkyl), —N(C₁₋₈ alkyl)₂, —NH((C₆)aryl), or —N((C₆)aryl)₂;formyl; ketones, such as —CO(C₁₋₈ alkyl), —CO((C₆aryl) esters, such as—CO₂(C₁₋₈ alkyl) and —CO₂ (C₆aryl). One of skill in art can readilychoose a suitable substituent based on the stability and pharmacologicaland synthetic activity of the compound of the present disclosure.

The term “pharmaceutically acceptable carrier” as used herein refers toany and all solvents, dispersion media, coatings, isotonic andabsorption delaying agents, and the like, that are compatible withpharmaceutical administration. The use of such media and agents forpharmaceutically active substances is well known in the art. Thecompositions may also contain other active compounds providingsupplemental, additional, or enhanced therapeutic functions.

The term “pharmaceutically acceptable composition” as used herein refersto a composition comprising at least one compound as disclosed hereinformulated together with one or more pharmaceutically acceptablecarriers.

The term “pharmaceutically acceptable prodrugs” as used hereinrepresents those prodrugs of the compounds of the present disclosurethat are, within the scope of sound medical judgment, suitable for usein contact with the tissues of humans and lower animals without unduetoxicity, irritation, allergic response, commensurate with a reasonablebenefit/risk ratio, and effective for their intended use, as well as thezwitterionic forms, where possible, of the compounds of the presentdisclosure. A discussion is provided in Higuchi et al., “Prodrugs asNovel Delivery Systems,” ACS Symposium Series, Vol. 14, and in Roche, E.B., ed. Bioreversible Carriers in Drug Design, American PharmaceuticalAssociation and Pergamon Press, 1987, both of which are incorporatedherein by reference.

The term “pharmaceutically acceptable salt(s)” refers to salts of acidicor basic groups that may be present in compounds used in the presentcompositions. Compounds included in the present compositions that arebasic in nature are capable of forming a wide variety of salts withvarious inorganic and organic acids. The acids that may be used toprepare pharmaceutically acceptable acid addition salts of such basiccompounds are those that form non-toxic acid addition salts, i.e., saltscontaining pharmacologically acceptable anions, including but notlimited to sulfate, citrate, matate, acetate, oxalate, chloride,bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate,isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate,tannate, pantothenate, bitartrate, ascorbate, succinate, maleate,gentisinate, fumarate, gluconate, glucaronate, saccharate, formate,benzoate, glutamate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate and pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Compounds includedin the present compositions that include an amino moiety may formpharmaceutically acceptable salts with various amino acids, in additionto the acids mentioned above. Compounds included in the presentcompositions, that are acidic in nature are capable of forming basesalts with various pharmacologically acceptable cations. Examples ofsuch salts include alkali metal or alkaline earth metal salts and,particularly, calcium, magnesium, sodium, lithium, zinc, potassium, andiron salts.

The compounds of the disclosure may contain one or more chiral centersand/or double bonds and, therefore, exist as stereoisomers, such asgeometric isomers, enantiomers or diastereomers. The term“stereoisomers” when used herein consist of all geometric isomers,enantiomers or diastereomers. These compounds may be designated by thesymbols “R” or “S,” depending on the configuration of substituentsaround the stereogenic carbon atom. The present disclosure encompassesvarious stereoisomers of these compounds and mixtures thereof.Stereoisomers include enantiomers and diastereomers. Mixtures ofenantiomers or diastereomers may be designated “(±)” in nomenclature,but the skilled artisan will recognize that a structure may denote achiral center implicitly.

Individual stereoisomers of compounds of the present disclosure can beprepared synthetically from commercially available starting materialsthat contain asymmetric or stereogenic centers, or by preparation ofracemic mixtures followed by resolution methods well known to those ofordinary skill in the art. These methods of resolution are exemplifiedby (1) attachment of a mixture of enantiomers to a chiral auxiliary,separation of the resulting mixture of diastereomers byrecrystallization or chromatography and liberation of the optically pureproduct from the auxiliary, (2) salt formation employing an opticallyactive resolving agent, or (3) direct separation of the mixture ofoptical enantiomers on chiral chromatographic columns. Stereoisomericmixtures can also be resolved into their component stereoisomers bywell-known methods, such as chiral-phase gas chromatography,chiral-phase high performance liquid chromatography, crystallizing thecompound as a chiral salt complex, or crystallizing the compound in achiral solvent. Stereoisomers can also be obtained fromstereomerically-pure intermediates, reagents, and catalysts bywell-known asymmetric synthetic methods.

Geometric isomers can also exist in the compounds of the presentdisclosure. The present disclosure encompasses the various geometricisomers and mixtures thereof resulting from the arrangement ofsubstituents around a carbon-carbon double bond or arrangement ofsubstituents around a carbocyclic ring. Substituents around acarbon-carbon double bond are designated as being in the “Z” or “E”configuration wherein the terms “Z” and “E” are used in accordance withIUPAC standards. Unless otherwise specified, structures depicting doublebonds encompass both the E and Z isomers.

Substituents around a carbon-carbon double bond alternatively can bereferred to as “cis” or “trans,” where “cis” represents substituents onthe same side of the double bond and “trans” represents substituents onopposite sides of the double bond. The arrangements of substituentsaround a carbocyclic ring are designated as “cis” or “trans.” The term“cis” represents substituents on the same side of the plane of the ringand the term “trans” represents substituents on opposite sides of theplane of the ring. Mixtures of compounds wherein the substituents aredisposed on both the same and opposite sides of plane of the ring aredesignated “cis/trans.”

The compounds disclosed herein may exist as tautomers and bothtautomeric forms are intended to be encompassed by the scope of thepresent disclosure, even though only one tautomeric structure isdepicted. For example, any claim to compound A below is understood toinclude tautomeric structure B, and vice versa, as well as mixturesthereof.

EXEMPLARY EMBODIMENTS

One embodiment of the invention provides a compound of Formula I:

or a stereoisomer, tautomer, pharmaceutical acceptable salt, or hydratethereof,wherein:

W₁ is selected from N and CR₁;

W₂ is selected from N and CR₂;

W₃ is selected from N and CR₃;

W₄ is selected from N and CR₄;

each W may be the same or different from each other;

A is selected from N and CH;

R₁, R₂, R₃, and R₄ are each independently selected from hydrogen, alkyl,alkenyl, alkynyl, alkoxy, thioalkyl, aryloxy, amino, aryl, hydroxyl, andhalogen, with the proviso that at least one of R₁-R₄ is not hydrogen;

two adjacent substituents selected from R₁, R₂, R₃, and R₄ may beconnected in a 5- or 6-membered ring to form a bicyclic carbocycle orbicyclic heterocycle;

AR1 is a group selected from the following:

B is a group selected from the following:

each ring system may be substituted with one or more substituentsindependently selected from R₁₀ and R₁₁;

R₅ is selected from hydrogen, alkoxy, alkyl, thioalkyl, aryloxy, aryl,hydroxyl, and halogen;

R₆ is selected from hydrogen, alkoxy, alkyl, thioalkyl, aryloxy, aryl,and halogen, with the proviso that if R₆=hydrogen, then R₂ is not —NMe₂;

R₇ is selected from hydrogen, alkyl, —SO₂R₁₂, —C(O)NR₁₂R₁₃, —C(O)R₁₂;

R₈ and R₉ are independently selected from hydrogen, aryl, alkenyl,alkyl, —SO₂R₁₂, —C(O)NR₁₂R₁₃, —C(O)R₁₂;

R₁₀ and R₁₁ are independently selected from hydrogen, halogen, alkyl,alkoxy, aryl, and hydroxyl;

R₁₂ and R₁₃ are independently selected from hydrogen, aryl, and alkyl;

Y is selected from NH, O, and S; and

two adjacent substituents selected from R₅, R₆, R₈, R₉, R₁₀, and R₁₁ maybe connected in a 5- or 6-membered ring to form a carbocycle orheterocycle.

In some embodiments, R₁, R₂, R₃, and R₄ are independently selected fromhydrogen, alkyl (such as methyl and ethyl), alkoxy (such as methoxy andethoxy), halogen (such as fluoride), and amino.

In some embodiments, R₁, R₂, R₃, and R₄ are each independently selectedfrom alkyl, alkoxy, or thioalkyl, each of which may be optionallysubstituted with hydroxyl, amino, halogen, or ester.

In some embodiments, R₁, R₂, R₃, and R₄ are each independently selectedfrom alkenyl or alkynyl, each of which may be optionally substitutedwith halogen.

In some embodiments, R₁, R₂, R₃, and R₄ are each independently selectedfrom aryl or aryloxy, each of which may be optionally substituted withhalogen, alkoxy, or amino.

In certain embodiments, R₁ and R₃ in the compound of Formula I areindependently selected from alkoxy (such as methoxy), halogen (such asfluoride), and amino (such as a substituted piperazine).

In other embodiments, R₁ and R₃ are independently selected from alkoxy.

In an exemplary compound of Formula I, R₁ and R₃ are methoxy and R₂ andR₄ are hydrogen.

In some embodiments R₅ is selected from hydrogen, alkyl (such as methyl,ethyl, propyl, and isopropyl), alkoxy (such as methoxy, ethoxy, propoxy,and isopropoxy, —OCF₃), halogen (such as fluoride and chloride).

In some embodiments R₅ is selected from alkoxy, alkyl, or thioalkyl,each of which may be optionally substituted with halogen, alkoxy, orhydroxyl.

some embodiments R₅ is selected from aryl or aryloxy, each of which maybe optionally substituted with halogen or alkoxy.

In other embodiments R₅ in compounds of Formula I is selected fromhydrogen, methoxy, ethoxy, —OCF₃, fluoride, chloride, methyl, and ethyl.

In exemplary embodiments R₆ in the compound of Formula I is selectedfrom hydrogen and alkoxy.

In other embodiments R₆ is selected from hydrogen, methoxy, ethoxy, oralkoxy optionally substituted with a hydroxyl or amino.

In some embodiments R₆ is selected from alkyl, alkoxy, or thioalkyl,each of which may be optionally substituted with halogen, amino,hydroxyl, or alkoxy.

some embodiments R₆ is selected from aryl or aryloxy, each of which maybe optionally substituted with halogen, alkoxy, or amino.

In certain embodiments R₆ is selected from hydrogen, methoxy,

In some embodiments Y is N.

In exemplary embodiments of compounds of Formula I, AR1 is selected from

In some embodiments, AR1 in the compound of Formula I is

In alternate embodiments, AR1 is

In certain exemplary compounds of Formula I, B is selected from

In some exemplary compounds of Formula I, B is selected from

In some embodiments, B is

In some embodiments R₇ in the compounds of Formula I is selected fromhydrogen and alkyl.

In some embodiments R₇ is alkyl optionally substituted with halogen,alkoxy, or amino.

In exemplary compounds of Formula I, R₈ and R₉ are independentlyselected from hydrogen and alkyl (such as methyl and ethyl).

In some embodiments R₈ and R₉ are each selected from aryl, alkyl, oralkenyl, each of which may be optionally substituted with halogen,hydroxyl, cyano, amido, sulfone, sulfonamide, heterocycle, or phosphate.

In exemplary embodiments R₁₀ and R₁₁ are independently selected fromhydrogen and halogen.

In some embodiments R₁₀ and R₁₁ are each selected from alkyl, alkoxy, oraryl, each of which may be optionally substituted with halogen.

In certain compounds of Formula I R₁₀ and R₁₁ are hydrogen.

In other exemplary embodiments, R₁₂ and R₁₃ are independently selectedfrom hydrogen and alkyl.

In some embodiments R₁₂ and R₁₃ are each selected from alkyl or aryl,each of which may be optionally substituted with halogen.

In other compounds of Formula I, R₁₂ and R₁₃ are independently selectedfrom hydrogen, methyl, ethyl, propyl, and isopropyl.

In certain embodiments of the invention, the compound of Formula I isselected from:

-   2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one    [Example 1],-   2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7-dimethoxyquinazolin-4(3H)-one    [Example 2],-   5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    [Example 3],-   2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    [Example 4],-   2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    [Example 5],-   5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    [Example 6],-   Methyl    2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetate    [Example 7],-   2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    [Example 8],-   2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanamide    [Example 9],-   2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetic    acid [Example 10],-   3-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoic    acid [Example 11],-   2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    bis(trifluoroacetate) [Example 12],-   2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    acetate [Example 13],-   2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    [Example 14],-   5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    Hydrochloride (Example 15),-   5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-one    Hydrochloride (Example 16),-   2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    Hydrochloride (Example 17),-   5,7-Dimethoxy-2-(6-(4-(methylsulfonyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    Hydrochloride (Example 18),-   5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    dihydrochloride (Example 19),-   2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one    (Example 20),-   7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one    Hydrochloride (Example 21),-   7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one    Hydrochloride (Example 22),-   7-(4-Isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one    Hydrochloride (Example 23),-   2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxy-7-phenylquinazolin-4(3H)-one    (Example 25),-   8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5-methoxyquinazolin-4(3H)-one    (Example 26),-   4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-isopropylpiperazine-1-carboxamide    Hydrochloride (Example 27),-   2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    Hydrochloride (Example 28),-   2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    (Example 29),-   5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    (Example 30),-   2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one    (Example 31),-   2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one    (Example 32).

In certain embodiments, the compound of Formula I is2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride (Example 17).

In selected embodiments of the compounds of Formula I, R₆ is selectedfrom the group represented by Formula II:

wherein:

D is selected from O and S;

E is selected from O, N, and S;

R₁₄ and R₁₅ are independently selected from hydrogen, alkyl, andcycloalkyl, and only one of R₁₄ and R₁₅ are present if E is O or S; and

n is selected from 1, 2, and 3. In some embodiments, D is oxygen. Insome embodiments, n=1. In some embodiments, R₁₄ and R₁₅ areindependently selected from hydrogen, and alkyl (such as C1-C5).

In some embodiments R₁₄ and R₁₅ are each selected from alkyl orcycloalkyl, each of which may be optionally substituted with halogen,amino, or hydroxyl.

In certain compounds of Formula I, R₆ is selected from hydrogen,methoxy,

Another aspect of the invention provides a method for inhibition of BETprotein function by binding to bromodomains, and their use in thetreatment and prevention of diseases and conditions in a mammal (e.g., ahuman) comprising administering a therapeutically effective amount of acompound of Formula I.

In one embodiment, because of potent effects of BET inhibitors in vitroon IL-6 and IL-17 transcription, BET inhibitor compounds of Formula Iare used as therapeutics for inflammatory disorders in which IL-6 and/orIL-17 have been implicated as drivers of disease. The followingautoimmune diseases are amenable to therapeutic use of BET inhibition byadministration of a compound of Formula I because of a prominent role ofIL-6 and/or IL-17: Acute Disseminated Encephalomyelitis [69],Agammaglobulinemia [70], Allergic Disease [71], Ankylosing spondylitis[72], Anti-GBM/Anti-TBM nephritis [73], Anti-phospholipid syndrome [74],Autoimmune aplastic anemia [75], Autoimmune hepatitis [76], Autoimmuneinner ear disease [77], Autoimmune myocarditis [78], Autoimmunepancreatitis [79], Autoimmune retinopathy [80], Autoimmunethrombocytopenic purpura [81], Behcet's Disease [82], Bullous pemphigoid[83], Castleman's Disease [84], Celiac Disease [85], Churg-Strausssyndrome [86], Crohn's Disease [87], Cogan's syndrome [88], Dry eyesyndrome [89], Essential mixed cryoglobulinemia [90], Dermatomyositis[91], Devic's Disease[92], Encephalitis [93], Eosinophlic esophagitis[94], Eosinophilic fasciitis [94], Erythema nodosum [95], Giant cellarteritis [96], Glomerulonephritis [97], Goodpasture's syndrome [73],Granulomatosis with Polyangiitis (Wegener's) [98], Graves' Disease[99],Guillain-Barre syndrome [100], Hashimoto's thyroiditis [101], Hemolyticanemia [102], Henoch-Schonlein purpura [103], IgA nephropathy [104],Inclusion body myositis [105], Type I diabetes [8], Interstitialcystitis [106], Kawasaki's Disease[107], Leukocytoclastic vasculitis[108], Lichen planus [109], Lupus (SLE) [110], Microscopic polyangitis[111], Multiple sclerosis [112], Myasthenia gravis [113], myositis [91],Optic neuritis [114], Pemphigus [115], POEMS syndrome [116],Polyarteritis nodosa [117], Primary biliary cirrhosis [118], Psoriasis[119], Psoriatic arthritis [120], Pyoderma gangrenosum [121], Relapsingpolychondritis [122], Rheumatoid arthritis [123], Sarcoidosis [124],Scleroderma [125], Sjogren's syndrome [126], Takayasu's arteritis [127],Transverse myelitis [128], Ulcerative colitis [129], Uveitis [130],Vitiligo [131].

Acute and chronic (non-autoimmune) inflammatory diseases characterizedby increased expression of pro-inflammatory cytokines, including IL-6,MCP-1, and IL-17, would also be amenable to therapeutic BET inhibition.These include, but are not limited to, sinusitis [132], pneumonitis[133], osteomyelitis [134], gastritis [135], enteritis [136], gingivitis[137], appendicitis [138], irritable bowel syndrome [139], tissue graftrejection [140], chronic obstructive pulmonary disease (COPD) [141],septic shock (toxic shock syndrome, SIRS, bacterial sepsis, etc) [12],osteoarthritis [142], acute gout [143], acute lung injury [141], acuterenal failure [144], burns [145], Herxheimer reaction [146], and SIRSassociated with viral infections [8].

In one embodiment, BET inhibitor compounds of Formula I are used fortreating rheumatoid arthritis (RA) and multiple sclerosis (MS). Strongproprietary data exist for the utility of BET inhibitors in preclinicalmodels of RA and MS [17]. Both RA and MS are characterized by adysregulation of the IL-6 and IL-17 inflammatory pathways [10] and thuswould be especially sensitive to BET inhibition. In another embodiment,BET inhibitor compounds of Formula I are used for treating sepsis andassociated afflictions. BET inhibition has been shown to inhibitdevelopment of sepsis, in part, by inhibiting IL-6 expression, inpreclinical models in both published [12] and proprietary data.

In one embodiment, BET inhibitor compounds of Formula I are used totreat cancer. Cancers that have an overexpression, translocation,amplification, or rearrangement c-myc or other myc family oncoproteins(MYCN, L-myc) are particularly sensitive to BET inhibition [27, 28].These cancers include, but are not limited to, B-acute lymphocyticleukemia, Burkitt's lymphoma, Diffuse large cell lymphoma, Multiplemyeloma, Primary plasma cell leukemia, Atypical carcinoid lung cancer,Bladder cancer, Breast cancer, Cervix cancer, Colon cancer, Gastriccancer, Glioblastoma, Hepatocellular carcinoma, Large cellneuroendocrine carcinoma, Medulloblastoma, Melanoma, nodular, Melanoma,superficial spreading, Neuroblastoma, esophageal squamous cellcarcinoma, Osteosarcoma, Ovarian cancer, Prostate cancer, Renal clearcell carcinoma, Retinoblastoma, Rhabdomyosarcoma, Small cell lungcarcinoma [25].

In one embodiment, BET inhibitor compounds of Formula I are used totreat cancers that result from an aberrant regulation (overexpression,translocation, etc) of BET proteins. These include, but are not limitedto, NUT midline carcinoma (Brd3 or Brd4 translocation to nutlin 1 gene)[22], B-cell lymphoma (Brd2 overexpression) [23], non-small cell lungcancer (BrdT overexpression) [147, 148], esophageal cancer and head andneck squamous cell carcinoma (BrdT overexpression) [147], colon cancer(Brd4) [149].

In one embodiment, because BET inhibitors decrease Brd-dependentrecruitment of pTEFb to genes involved in cell proliferation, BETinhibitor compounds of Formula I are used to treat cancers that rely onpTEFb (Cdk9/cyclin T) and BET proteins to regulate oncogenes. Theseinclude, but are not limited to, chronic lymphocytic leukemia andmultiple myeloma [150], follicular lymphoma, diffuse large B celllymphoma with germinal center phenotype, Burkitt's lymphoma, Hodgkin'slymphoma, follicular lymphomas and activated, anaplastic large celllymphoma [151], neuroblastoma and primary neuroectodermal tumor [152],rhabdomyosarcoma [153], prostate cancer [154], and breast cancer [45].

In one embodiment, BET inhibitor compounds of Formula I are used totreat cancers in which BET-responsive genes, such as CDK6, Bcl2, TYRO3,MYB, and hTERT are up-regulated [26, 27]. These cancers include, but arenot limited to, pancreatic cancer, breast cancer, colon cancer,glioblastoma, adenoid cystic carcinoma, T-cell prolymphocytic leukemia,malignant glioma, bladder cancer, medulloblastoma, thyroid cancer,melanoma, multiple myeloma, Barret's adenocarcinoma, hepatoma, prostatecancer, pro-myelocytic leukemia, chronic lymphocytic leukemia, mantlecell lymphoma, diffuse large B-cell lymphoma, small cell lung cancer,renal carcinoma [32, 155-162].

Published and proprietary data have shown direct effects of BETinhibition on cell proliferation in various cancers. In one embodiment,BET inhibitor compounds of Formula I are used to treat cancers for whichexist published and, for some, proprietary, in vivo and/or in vitro datashowing a direct effect of BET inhibition on cell proliferation. Thesecancers include NMC (NUT-midline carcinoma), acute myeloid leukemia(AML), acute B lymphoblastic leukemia (B-ALL), Burkitt's Lymphoma,B-cell Lymphoma, Melanoma, mixed lineage leukemia, multiple myeloma,pro-myelocytic leukemia (PML), non-Hodgkin's lymphoma [24, 26-30, 33].Examples provided within this application have also shown a directeffect of BET inhibition on cell proliferation in vitro for thefollowing cancers: Neuroblastoma, Medulloblastoma, lung carcinoma(NSCLC, SCLC), and colon carcinoma.

In one embodiment, because of potential synergy or additive effectsbetween BET inhibitors and other cancer therapy, BET inhibitor compoundsof Formula I are combined with other therapies, chemotherapeutic agents,or anti-proliferative agents to treat human cancer and otherproliferative disorders. The list of therapeutic agents which can becombined with BET inhibitors in cancer treatment includes, but is notlimited to, ABT-737, Azacitidine (Vidaza), AZD1152 (Barasertib), AZD2281(Olaparib), AZD6244 (Selumetinib), BEZ235, Bleomycin Sulfate, Bortezomib(Velcade), Busulfan (Myleran), Camptothecin, Cisplatin, Cyclophosphamide(Clafen), CYT387, Cytarabine (Ara-C), Dacarbazine, DAPT (GSI-IX),Decitabine, Dexamethasone, Doxorubicin (Adriamycin), Etoposide,Everolimrus (RAD001), Flavopiridol (Alvocidib), Ganetespib (STA-9090),Gefitinib (Iressa), Idarubicin, Ifosfamide (Mitoxana), IFNa2a (RoferonA), Melphalan (Alkeran), Methazolastone (temozolomide), Metformin,Mitoxantrone (Novantrone), Paclitaxel, Phenformin, PKC412 (Midostaurin),PLX4032 (Vemurafenib), Pomalidomide (CC-4047), Prednisone (Deltasone),Rapamycin, Revlimid (Lenalidomide), Ruxolitinib (INCB018424), Sorafenib(Nexavar), SU11248 (Sunitinib), SU11274, Vinblastine, Vincristine(Oncovin), Vinorelbine (Navelbine), Vorinostat (SAHA), WP1130(Degrasyn).

In one embodiment, because of their ability to up-regulate ApoA-1transcription and protein expression [11, 35], BET inhibitor compoundsof Formula I are used to treat cardiovascular diseases that aregenerally associated with including dyslipidemia, atherosclerosis,hypercholesterolemia, and metabolic syndrome [8, 19]. In anotherembodiment, BET inhibitor compounds of Formula I are used to treatnon-cardiovascular disease characterized by deficits in ApoA-1,including Alzheimer's disease [37].

In one embodiment, BET inhibitor compounds of Formula I are used inpatients with insulin resistance and type II diabetes [8, 19, 38, 39].The anti-inflammatory effects of BET inhibition would have additionalvalue in decreasing inflammation associated with diabetes and metabolicdisease [163].

In one embodiment, because of their ability to down-regulate viralpromoters, BET inhibitor compounds of Formula I are used as therapeuticsfor cancers that are associated with viruses including Epstein-BarrVirus (EBV), hepatitis virus (HBV, HCV), Kaposi's sarcoma associatedvirus (KSHV), human papilloma virus (HPV), Merkel cell polyomavirus, andhuman cytomegalovirus (CMV) [40-42, 164]. In another embodiment, becauseof their ability to reactivate HIV-1 in models of latent T cellinfection and latent monocyte infection, BET inhibitors could be used incombination with anti-retroviral therapeutics for treating HIV [43-46].

In one embodiment, because of the role of epigenetic processes andbromodomain-containing proteins in neurological disorders, BET inhibitorcompounds of Formula I are used to treat diseases including, but notlimited to, Alzheimer's disease, Parkinson's disease, Huntingtondisease, bipolar disorder, schizophrenia, Rubinstein-Taybi syndrome, andepilepsy [9, 165].

In one embodiment, because of the effect of BRDT depletion or inhibitionon spermatid development, BET inhibitor compounds of Formula I are usedas reversible, male contraceptive agents [50, 51].

Pharmaceutical Compositions

Pharmaceutical compositions of the present disclosure comprise at leastone compound of Formula I, or tautomer, stereoisomer, pharmaceuticallyacceptable salt or hydrate thereof formulated together with one or morepharmaceutically acceptable carriers. These formulations include thosesuitable for oral, rectal, topical, buccal and parenteral (e.g.,subcutaneous, intramuscular, intradermal, or intravenous)administration. The most suitable form of administration in any givencase will depend on the degree and severity of the condition beingtreated and on the nature of the particular compound being used.

Formulations suitable for oral administration may be presented indiscrete units, such as capsules, cachets, lozenges, or tablets, eachcontaining a predetermined amount of a compound of the presentdisclosure as powder or granules; as a solution or a suspension in anaqueous or non-aqueous liquid; or as an oil-in-water or water-in-oilemulsion. As indicated, such formulations may be prepared by anysuitable method of pharmacy which includes the step of bringing intoassociation at least one compound of the present disclosure as theactive compound and a carrier or excipient (which may constitute one ormore accessory ingredients). The carrier must be acceptable in the senseof being compatible with the other ingredients of the formulation andmust not be deleterious to the recipient. The carrier may be a solid ora liquid, or both, and may be formulated with at least one compounddescribed herein as the active compound in a unit-dose formulation, forexample, a tablet, which may contain from about 0.05% to about 95% byweight of the at least one active compound. Other pharmacologicallyactive substances may also be present including other compounds. Theformulations of the present disclosure may be prepared by any of thewell-known techniques of pharmacy consisting essentially of admixing thecomponents.

For solid compositions, conventional nontoxic solid carriers include,for example, pharmaceutical grades of mannitol, lactose, starch,magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose,magnesium carbonate, and the like. Liquid pharmacologicallyadministrable compositions can, for example, be prepared by, forexample, dissolving or dispersing, at least one active compound of thepresent disclosure as described herein and optional pharmaceuticaladjuvants in an excipient, such as, for example, water, saline, aqueousdextrose, glycerol, ethanol, and the like, to thereby form a solution orsuspension. In general, suitable formulations may be prepared byuniformly and intimately admixing the at least one active compound ofthe present disclosure with a liquid or finely divided solid carrier, orboth, and then, if necessary, shaping the product. For example, a tabletmay be prepared by compressing or molding a powder or granules of atleast one compound of the present disclosure, which may be optionallycombined with one or more accessory ingredients. Compressed tablets maybe prepared by compressing, in a suitable machine, at least one compoundof the present disclosure in a free-flowing form, such as a powder orgranules, which may be optionally mixed with a binder, lubricant, inertdiluent and/or surface active/dispersing agent(s). Molded tablets may bemade by molding, in a suitable machine, where the powdered form of atleast one compound of the present disclosure is moistened with an inertliquid diluent.

Formulations suitable for buccal (sub-lingual) administration includelozenges comprising at least one compound of the present disclosure in aflavored base, usually sucrose and acacia or tragacanth, and pastillescomprising the at least one compound in an inert base such as gelatinand glycerin or sucrose and acacia.

Formulations of the present disclosure suitable for parenteraladministration comprise sterile aqueous preparations of at least onecompound of Formula I or tautomers, stereoisomers, pharmaceuticallyacceptable salts, and hydrates thereof, which are approximately isotonicwith the blood of the intended recipient. These preparations areadministered intravenously, although administration may also be effectedby means of subcutaneous, intramuscular, or intradermal injection. Suchpreparations may conveniently be prepared by admixing at least onecompound described herein with water and rendering the resultingsolution sterile and isotonic with the blood. Injectable compositionsaccording to the present disclosure may contain from about 0.1 to about5% w/w of the active compound.

Formulations suitable for rectal administration are presented asunit-dose suppositories. These may be prepared by admixing at least onecompound as described herein with one or more conventional solidcarriers, for example, cocoa butter, and then shaping the resultingmixture.

Formulations suitable for topical application to the skin may take theform of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil.Carriers and excipients which may be used include Vaseline, lanoline,polyethylene glycols, alcohols, and combinations of two or more thereof.The active compound (i.e., at least one compound of Formula I ortautomers, stereoisomers, pharmaceutically acceptable salts, andhydrates thereof) is generally present at a concentration of from about0.1% to about 15% w/w of the composition, for example, from about 0.5 toabout 2%.

The amount of active compound administered may be dependent on thesubject being treated, the subject's weight, the manner ofadministration and the judgment of the prescribing physician. Forexample, a dosing schedule may involve the daily or semi-dailyadministration of the encapsulated compound at a perceived dosage ofabout 1 μg to about 1000 mg. In another embodiment, intermittentadministration, such as on a monthly or yearly basis, of a dose of theencapsulated compound may be employed. Encapsulation facilitates accessto the site of action and allows the administration of the activeingredients simultaneously, in theory producing a synergistic effect. Inaccordance with standard dosing regimens, physicians will readilydetermine optimum dosages and will be able to readily modifyadministration to achieve such dosages.

A therapeutically effective amount of a compound or compositiondisclosed herein can be measured by the therapeutic effectiveness of thecompound. The dosages, however, may be varied depending upon therequirements of the patient, the severity of the condition beingtreated, and the compound being used. In one embodiment, thetherapeutically effective amount of a disclosed compound is sufficientto establish a maximal plasma concentration. Preliminary doses as, forexample, determined according to animal tests, and the scaling ofdosages for human administration is performed according to art-acceptedpractices.

Toxicity and therapeutic efficacy can be determined by standardpharmaceutical procedures in cell cultures or experimental animals,e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀.Compositions that exhibit large therapeutic indices are preferable.

Data obtained from the cell culture assays or animal studies can be usedin formulating a range of dosage for use in humans. Therapeuticallyeffective dosages achieved in one animal model may be converted for usein another animal, including humans, using conversion factors known inthe art (see, e.g., Freireich et al., Cancer Chemother. Reports50(4):219-244 (1966) and the following Table for Equivalent Surface AreaDosage Factors).

Equivalent Surface Area Dosage Factors:

To: Mouse Rat Monkey Dog Human From: (20 g) (150 g) (3.5 kg) (8 kg) (60kg) Mouse  1 1/2 1/4 1/6 1/12 Rat  2 1 1/2 1/4 1/7  Monkey  4 2 1 3/51/3  Dog  6 4 3/5 1 1/2  Human 12 7 3 2 1

The dosage of such compounds lies preferably within a range ofcirculating concentrations that include the ED₅₀ with little or notoxicity. The dosage may vary within this range depending upon thedosage form employed and the route of administration utilized.Generally, a therapeutically effective amount may vary with thesubject's age, condition, and gender, as well as the severity of themedical condition in the subject. The dosage may be determined by aphysician and adjusted, as necessary, to suit observed effects of thetreatment.

In one embodiment, a compound of Formula I or a tautomer, stereoisomer,pharmaceutically acceptable salt or hydrate thereof, is administered incombination with another therapeutic agent. The other therapeutic agentcan provide additive or synergistic value relative to the administrationof a compound of the present disclosure alone. The therapeutic agent canbe, for example, a statin; a PPAR agonist, e.g., a thiazolidinedione orfibrate; a niacin, a RVX, FXR or LXR agonist; a bile-acid reuptakeinhibitor; a cholesterol absorption inhibitor; a cholesterol synthesisinhibitor; a cholesteryl ester transfer protein (CETP), an ion-exchangeresin; an antioxidant; an inhibitor of AcylCoA cholesterolacyltransferase (ACAT inhibitor); a tyrophostine; a sulfonylurea-baseddrug; a biguanide; an alpha-glucosidase inhibitor; an apolipoprotein Eregulator; a HMG-CoA reductase inhibitor, a microsomal triglyceridetransfer protein; an LDL-lowing drug; an HDL-raising drug; an HDLenhancer; a regulator of the apolipoprotein A-IV and/or apolipoproteingenes; or any cardiovascular drug.

In another embodiment, a compound of Formula I or a tautomer,stereoisomer, pharmaceutically acceptable salt or hydrate thereof, isadministered in combination with one or more anti-inflammatory agents.Anti-inflammatory agents can include immunosuppressants, TNF inhibitors,corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs),disease-modifying anti-rheumatic drugs (DMARDS), and the like. Exemplaryanti-inflammatory agents include, for example, prednisone;methylprenisolone (Medrol®), triamcinolone, methotrexate (Rheumatrex®,Trexall®), hydroxychloroquine (Plaquenil®), sulfasalzine (Azulfidine®),leflunomide (Arava®), etanercept (Enbrel®), infliximab (Remicade®),adalimumab (Humira®), rituximab (Rituxan®), abatacept (Orencia®),interleukin-1, anakinra (Kineret™), ibuprofen, ketoprofen, fenoprofen,naproxen, aspirin, acetominophen, indomethacin, sulindac, meloxicam,piroxicam, tenoxicam, lornoxicam, ketorolac, etodolac, mefenamic acid,meclofenamic acid, flufenamic acid, tolfenamic acid, diclofenac,oxaprozin, apazone, nimesulide, nabumetone, tenidap, etanercept,tolmetin, phenylbutazone, oxyphenbutazone, diflunisal, salsalate,olsalazine, or sulfasalazine.

Specific embodiments of the invention include:

1. A compound of Formula I:

or a stereoisomer, tautomer, pharmaceutical acceptable salt, or hydratethereof,wherein:

W₁ is selected from N and CR₁;

W₂ is selected from N and CR₂;

W₃ is selected from N and CR₃;

W₄ is selected from N and CR₄;

each W may be the same or different from each other;

A is selected from N and CH;

R₁, R₂, R₃, and R₄ are each independently selected from hydrogen, alkyl,alkenyl, alkynyl, alkoxy, thioalkyl, aryloxy, aryl, amino, hydroxyl, andhalogen;

two adjacent substituents selected from R₁, R₂, R₃, and R₄ may beconnected in a 5- or 6-membered ring to form a bicyclic carbocycle orbicyclic heterocycle;

AR1 is a group selected from the following:

B is a group selected from the following:

each ring system may be substituted with one or more substituentsindependently selected from R₁₀ and R₁₁;

R₅ is selected from hydrogen, alkoxy, alkyl, thioalkyl, aryloxy, aryl,hydroxyl, and halogen;

R₆ is selected from hydrogen, alkoxy, alkyl, thioalkyl, aryloxy, aryl,and halogen;

R₇ is selected from hydrogen, alkyl, —SO₂R₁₂, —C(O)NR₁₂R₁₃, —C(O)R₁₂;

R₈ and R₉ are independently selected from hydrogen, aryl, alkenyl,alkyl, —SO₂R₁₂, —C(O)NR₁₂R₁₃, —C(O)R₁₂;

R₁₀ and R₁₁ are independently selected from hydrogen, halogen, alkyl,alkoxy, aryl, and hydroxyl;

R₁₂ and R₁₃ are independently selected from hydrogen, aryl, and alkyl;

Y is selected from NH, O, and S; and

two adjacent substituents selected from R₅, R₆, R₈, R₉, R₁₀, and R₁₁ maybe connected in a 5- or 6-membered ring to form a carbocycle orheterocycle.

2. The compound according to embodiment 1 wherein if R₆ is hydrogen,then R₂ is not —NMe₂.

3. The compound according to embodiment 1 wherein R₁, R₂, R₃, and R₄ areindependently selected from hydrogen, alkyl, alkoxy, halogen, and amino.

4. The compound according to embodiment 1, wherein at least one of R₁-R₄is not hydrogen.

5. The compound according to embodiment 1 wherein R₁ and R₃ areindependently selected from alkoxy, halogen, and amino.

6. The compound according to embodiment 1 wherein R₁ and R₃ areindependently selected methoxy, fluoride, and a substituted piperazine.

7. The compound according to embodiment 1 wherein R₁ and R₃ areindependently selected from alkoxy.

8. The compound according to embodiment 1 wherein R₁ and R₃ are methoxyand R₂ and R₄ are hydrogen.

9. The compound according to any one of embodiments 1-8 wherein R₅ isselected from hydrogen, alkyl, alkoxy, and halogen.

10. The compound according to embodiment 9 wherein R₅ is selected fromhydrogen, methyl, ethyl propyl, and isopropyl, methoxy, ethoxy, propoxy,isopropoxy, —OCF₃, fluoride, and chloride.

11. The compound according to any one of embodiments 1-8 wherein R₅ isselected from hydrogen, methoxy, ethoxy, —OCF₃, fluoride, chloride,methyl, and ethyl.

12. The compound according to any one of embodiments 1-11 wherein R₆ isselected from hydrogen and alkoxy optionally substituted with a hydroxylor amino.

13. The compound according to any one of embodiments 1-11 wherein R₆ isselected from hydrogen, methoxy, and ethoxy.

14. The compound according to any one of embodiments 1-11 wherein R₆ isselected from hydrogen, methoxy,

15. The compound according to any one of embodiments 1-14 wherein Y isN.

16. The compound according to any one of embodiments 1-15 wherein AR1 isselected from

17. The compound according to any one of embodiments 1-16 wherein AR1 is

18. The compound according to any one of embodiments 1-16 wherein AR1 is

19. The compound according to any one of embodiments 1-18 wherein B isselected from

20. The compound according to any one of embodiments 1-18 wherein B isselected from

21. The compound according to embodiment 20 wherein B is

22. The compound according to any one of embodiments 1-21, wherein R₇ isselected from hydrogen and alkyl.

23. The compound according to any one of embodiments 1-22, wherein R₈and R₉ are independently selected from hydrogen and alkyl.

24. The compound according to any one of embodiments 1-22, wherein R₈and R₉ are independently selected from hydrogen, methyl, and ethyl.

25. The compound according to any one of embodiments 1-24, wherein R₁₀and R₁₁ are independently selected from hydrogen and halogen.

26. The compound according to embodiment 25 wherein R₁₀ and R₁₁ arehydrogen.

27. The compound according to any one of embodiments 1-26 wherein R₁₂and R₁₃ are independently selected from hydrogen and alkyl.

28. The compound according to any one of embodiments 1-26 wherein R₁₂and R₁₃ are independently selected from hydrogen, methyl, ethyl, propyl,and isopropyl.

29. A compound according to embodiment 1 selected from:

-   2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;-   2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7-dimethoxyquinazolin-4(3H)-one;-   5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;-   2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;-   2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;-   5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;-   Methyl    2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetate;-   2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;-   2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanamide;-   2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetic    acid;-   3-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoic    acid;-   2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    bis(trifluoroacetate);-   2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    acetate;-   2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;-   5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    Hydrochloride;-   5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-one    Hydrochloride;-   2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    Hydrochloride;-   5,7-Dimethoxy-2-(6-(4-(methylsulfonyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    Hydrochloride;-   5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one    dihydrochloride;-   2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;-   7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one    Hydrochloride;-   7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one    Hydrochloride;-   7-(4-Isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one    Hydrochloride;-   2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxy-7-phenylquinazolin-4(3H)-one;-   8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5-methoxyquinazolin-4(3H)-one;-   4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-isopropylpiperazine-1-carboxamide    Hydrochloride;-   2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one    Hydrochloride;-   2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;-   5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;-   2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;-   2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one.

30. A compound of embodiment 1, wherein the compound is2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride.

31. A compound according to any one of embodiments 1-11 and 15-28wherein R₆ is selected from the group represented by Formula II:

wherein:

D is selected from O and S;

E is selected from O, N, and S;

R₁₄ and R₁₅ are independently selected from hydrogen, alkyl, andcycloalkyl, wherein if E is O or S, only one of R₁₄ and R₁₅ is present;and

n is selected from 1, 2, and 3.

32. The compound according to embodiment 31 wherein D is oxygen.

33. The compound according to embodiment 31 or embodiment 32 whereinn=1.

34. The compound according to any one of embodiments 31-33 wherein R₁₄and R₁₅ are independently selected from hydrogen, and alkyl.

35. The compound according to embodiment 34, wherein R₁₄ and R₁₅ areindependently selected from hydrogen, and C1-C5 alkyl.

36. The compound according to any one of embodiments 31-35 wherein R₆ isselected from hydrogen, methoxy,

, and

37. A pharmaceutical composition comprising a compound according to anyone of embodiments 1-36.

38. A compound according to any one of embodiments 1-36 for use as amedicament.

39. A method for inhibiting BET proteins in a mammal comprisingadministering a therapeutically effective amount of a compound accordingto any one of embodiments 1-36.

40. A method for treating a disease that is sensitive to a BET inhibitorcomprising administering a therapeutically effective amount of acompound according to any one of embodiments 1-36.

41. A method for treating an autoimmune disease in a mammal comprisingadministering a therapeutically effective amount of a compound accordingto any one of embodiments 1-36.

42. The method of embodiment 41, wherein the autoimmune disease isselected from Acute Disseminated Encephalomyelitis, Agammaglobulinemia,Allergic Disease, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis,Anti-phospholipid syndrome, Autoimmune aplastic anemia, Autoimmunehepatitis, Autoimmune inner ear disease, Autoimmune myocarditis,Autoimmune pancreatitis, Autoimmune retinopathy, Autoimmunethrombocytopenic purpura, Behcet's Disease, Bullous pemphigoid,Castleman's Disease, Celiac Disease, Churg-Strauss syndrome, Crohn'sDisease, Cogan's syndrome, Dry eye syndrome, Essential mixedcryoglobulinemia, Dermatomyositis, Devic's Disease, Encephalitis,Eosinophlic esophagitis, Eosinophilic fasciitis, Erythema nodosum, Giantcell arteritis, Glomerulonephritis, Goodpasture's syndrome,Granulomatosis with Polyangiitis (Wegener's), Graves' Disease,Guillain-Barre syndrome, Hashimoto's thyroiditis, Hemolytic anemia,Henoch-Schonlein purpura, IgA nephropathy, Inclusion body myositis, TypeI diabetes, Interstitial cystitis, Kawasaki's Disease, Leukocytoclasticvasculitis, Lichen planus, Lupus (SLE), Microscopic polyangitis,Multiple sclerosis, Myasthenia gravis, myositis, Optic neuritis,Pemphigus, POEMS syndrome, Polyarteritis nodosa, Primary biliarycirrhosis, Psoriasis, Psoriatic arthritis, Pyoderma gangrenosum,Relapsing polychondritis, Rheumatoid arthritis, Sarcoidosis,Scleroderma, Sjogren's syndrome, Takayasu's arteritis, Transversemyelitis, Ulcerative colitis, Uveitis, and Vitiligo.

43. A method for treating inflammatory diseases or disorders in a mammalcomprising administering a therapeutically effective amount of acompound according to any one of embodiments 1-36.

44. The method of embodiment 43 wherein the inflammatory disease ordisorder is selected from sinusitis, pneumonitis, osteomyelitis,gastritis, enteritis, gingivitis, appendicitis, irritable bowelsyndrome, tissue graft rejection, chronic obstructive pulmonary disease(COPD), septic shock, toxic shock syndrome, SIRS, bacterial sepsis,osteoarthritis, acute gout, acute lung injury, acute renal failure,burns, Herxheimer reaction, and SIRS associated with viral infections.

45. A method for treating or preventing a cancer in a mammal comprisingadministering a therapeutically effective amount of a compound accordingto any one of embodiments 1-36.

46. The method of embodiment 45 wherein the cancer is a midlinecarcinoma.

47. The method of embodiment 45 wherein the cancer exhibitsoverexpression, translocation, amplification, or rearrangement of a mycfamily oncoproteins.

48. The method of embodiment 45 wherein the cancer is characterized byoverexpression of c-myc.

49. The method of embodiment 45 wherein the cancer is characterized byis characterized by overexpression n-myc.

50. The method of embodiment 45 wherein the cancer results from aberrantregulation of BET proteins.

51. The method of embodiment 45 wherein the cancer is characterized byrecruitment of pTEFb to regulate oncogenes.

52. The method of embodiment 45 wherein the cancer is characterized byupregulation of CDK6, Bcl2, TYRO3, MYB and/or hTERT.

53. The method of embodiment 45 wherein the cancer is selected from:B-acute lymphocytic leukemia, Burkitt's lymphoma, diffuse large celllymphoma, multiple myeloma, primary plasma cell leukemia, atypicalcarcinoid lung cancer, bladder cancer, breast cancer, cervix cancer,colon cancer, gastric cancer, glioblastoma, hepatocellular carcinoma,large cell neuroendocrine carcinoma, medulloblastoma, melanoma, nodularmelanoma, neuroblastoma, oesophageal squamous cell carcinoma,osteosarcoma, ovarian cancer, prostate cancer, renal clear cellcarcinoma, retinoblastoma, rhabdomyosarcoma, small cell lung carcinoma,NUT midline carcinoma, B-cell lymphoma, non-small cell lung cancer,esophageal cancer and head and neck squamous cell carcinoma, chroniclymphocytic leukemia, follicular lymphoma, diffuse large B cell lymphomawith germinal center phenotype, Burkitt's lymphoma, Hodgkin's lymphoma,follicular lymphomas, activated anaplastic large cell lymphoma, primaryneuroectodermal tumor, pancreatic cancer, adenoid cystic carcinoma,T-cell prolymphocytic leukemia, malignant glioma, thyroid cancer,Barret's adenocarcinoma, hepatoma, pro-myelocytic leukemia, chroniclymphocytic leukemia, and mantle cell lymphoma.

54. The method of any one of embodiments 45-53 wherein the compound ofFormula I is administered in combination with another anticancer agent.

55. The method of embodiment 54, wherein the anticancer agent isselected from ABT-737, Azacitidine (Vidaza), AZD1152 (Barasertib),AZD2281 (Olaparib), AZD6244 (Selumetinib), BEZ235, Bleomycin Sulfate,Bortezomib (Velcade), Busulfan (Myleran), Camptothecin, Cisplatin,Cyclophosphamide (Clafen), CYT387, Cytarabine (Ara-C), Dacarbazine, DAPT(GSI-IX), Decitabine, Dexamethasone, Doxorubicin (Adriamycin),Etoposide, Everolimus (RAD001), Flavopiridol (Alvocidib), Ganetespib(STA-9090), Gefitinib (Iressa), Idarubicin, Ifosfamide (Mitoxana),IFNa2a (Roferon A), Melphalan (Alkeran), Methazolastone (temozolomide),Metformin, Mitoxantrone (Novantrone), Paclitaxel, Phenformin, PKC412(Midostaurin), PLX4032 (Vemurafenib), Pomalidomide (CC-4047), Prednisone(Deltasone), Rapamycin, Revlimid (Lenalidomide), Ruxolitinib(INCB018424), Sorafenib (Nexavar), SU11248 (Sunitinib), SU11274,Vinblastine, Vincristine (Oncovin), Vinorelbine (Navelbine), Vorinostat(SAHA), and WP1130 (Degrasyn).

56. A method of treating a cardiovascular disease comprisingadministering a therapeutically effective amount of a compound accordingto any one of embodiments 1-36.

57. The method of embodiment 56, wherein the cardiovascular disease isdyslipidemia, atherosclerosis, hypercholesterolemia, or metabolicsyndrome.

58. A method of treating insulin resistance diabetes comprisingadministering a therapeutically effective amount of a compound accordingto any one of embodiments 1-36.

59. A method of treating a neurological disorder comprisingadministering a therapeutically effective amount of a compound accordingto any one of embodiments 1-36.

59. The method of embodiment 58 wherein the neurological disorder isAlzheimer's disease, Parkinson's disease, Huntington disease, bipolardisorder, schizophrenia, Rubinstein-Taybi syndrome, or epilepsy.

60. A method of male contraception comprising administering atherapeutically effective amount of a compound according to any one ofembodiments 1-35.

61. A method of treating HIV comprising administering a therapeuticallyeffective amount of a compound according to any one of embodiments 1-35.

62. A method of treating a cancer associated with a viral infectioncomprising administering a therapeutically effective amount of acompound according to any one of embodiments 1-35.

63. The method of embodiment 62 wherein the virus is selected fromEpstein-Barr Virus, hepatitis B virus, hepatitis C virus, Kaposi'ssarcoma associated virus, human papilloma virus, Merkel cellpolyomavirus, and human cytomegalovirus.

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EXAMPLES Example 12-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one

To a solution of 2-amino-4,6-dimethoxy-benzamide (0.5 g, 2.55 mmol) inN,N-dimethylacetamide (30 mL) was added 3-bromo-benzaldehyde (0.32 mL,2.80 mmol) followed by NaHSO₃ (0.39 g, 3.82 mmol) and p-toluenesulfonicacid monohydrate (0.24 g, 1.27 mmol). The reaction was heated at 120° C.for 20 h. After that time it was cooled to room temperature (rt),concentrated under reduced pressure and diluted with water. Theprecipitated solids were collected by filtration, washed with water anddried under vacuum. The product was triturated with diethyl ether togive 2-(3-bromo-phenyl)-5,7-dimethoxy-3H-quinazolin-4-one (0.81 g, 88%)as a yellow solid: ¹H NMR (400 MHz, CDCl₃) δ 8.23 (s, 1H), 7.84 (d,J=7.0 Hz, 1H), 7.66 (d, J=7.0 Hz, 1H), 7.38 (t, J=7.0 Hz, 1H), 6.8 (s,1H), 6.46 (s, 1H), 3.94 (s, 3H), 3.90 (s, 3H).

A mixture of 2-(3-bromo-phenyl)-5,7-dimethoxy-3H-quinazolin-4-one (0.5g, 1.38 mmol), 1-isopropyl-piperazine (0.24 mL, 1.66 mmol), potassiumt-butoxide (0.32 g, 3.31 mmol),2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (60.1 mg, 0.096 mmol) andtris(dibenzylideneacetone)dipalladium(0) (31.6 mg, 0.034 mmol) innitrogen saturated toluene (10 mL) was heated at 100° C. for 48 h. Afterthat time the reaction was cooled to rt and concentrated under reducedpressure. The product was purified by flash column chromatography(silica gel, dichloromethane to 97:3 dichloromethane/methanol) to give2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one(0.080 g, 14%) as a yellow solid: mp 251-253° C.; ¹H NMR (400 MHz,CDCl₃): δ 7.56 (s, 1H), 7.42-7.38 (m, 2H), 7.13-7.09 (m, 1H), 6.84 (d,J=2.4 Hz, 1H), 6.46 (d, J=2.4 Hz, 1H), 3.98 (s, 3H), 3.94 (s, 3H),3.38-3.28 (m, 4H), 2.78-2.68 (m, 4H), 1.11 (d, 6H); ESI MS m/z 407(M−1)⁻

Example 22-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7-dimethoxyquinazolin-4(3H)-one

A solution of 1,3-dibromo-5-methoxybenzene (5.00 g, 18.8 mmol) inanhydrous diethyl ether (100 mL) was cooled to −78° C. Then a solutionof n-butyllithium in hexanes (2.5 M, 8.3 mL, 20.68 mmol) was addeddrop-wise at −78° C. under nitrogen. After the addition was complete,the reaction was stirred at −78° C. for 45 min. After that time,anhydrous DMF (7.3 mL), 94.0 mmol) was added, the cooling bath wasremoved and the reaction mixture was allowed to warm to rt. The reactionwas diluted with saturated aqueous NH₄Cl solution (100 mL) and diethylether (100 mL). The organic phase was separated, washed with water,dried over anhydrous Na₂SO₄ and concentrated under reduced pressure togive 3-bromo-5-methoxybenzaldehyde (3.95 g, 98%) as yellow oil: ¹H NMR(400 MHz, CDCl₃) δ 9.90 (s, 1H), 7.57-7.60 (m, 1H), 7.31-7.33 (m, 2H),3.86 (s, 3H).

A solution of 3-bromo-5-methoxybenzaldehyde (3.93 g, 18.2 mmol),propane-1,3-diol (1.6 mL, 21.8 mmol) and p-toluenesulfonic acidmonohydrate (0.104 g, 0.54 mmol) in anhydrous toluene (100 mL) wasbrought to reflux for 18 h under nitrogen. During this time water wasremoved using a Dean-Stark apparatus. After this time the reaction wascooled to rt, washed with 5% aqueous Na₂CO₃ solution (50 mL) and driedover anhydrous Na₂SO₄. The reaction was concentrated under reducedpressure to give 2-(3-bromo-5-methoxyphenyl)-1,3-dioxane (4.80 g, 96%)as a brown oil.

A mixture of 2-(3-bromo-5-methoxyphenyl)-1,3-dioxane (4.75 g, 17.4mmol), 1-isopropyl-piperazine (3.14 mL, 21.96 mmol),sodium-tert-butoxide (3.00 g, 31.3 mmol),2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (0.342 g, 0.55 mmol) andtris(dibenzylideneacetone)dipalladium (0) (0.167 g, 0.18 mmol) intoluene (40 mL) was stirred at 100° C. for 18 h. After this time thereaction was cooled to rt and the dark brown mixture was poured into anice-cold solution of 1N HCl (100 mL) and stirred vigorously for 2 h atrt. After this time the reaction mixture was re-cooled to 0° C. and thepH was adjusted to 13 with 6N NaOH solution. The reaction mixture wasthen extracted with diethyl ether. The organic phase was concentrated,taken up in 2N HCl (50 mL) and stirred for 1 h. The resulting mixturewas extracted with dichloromethane (2×150 mL). The pH of the aqueousphase was adjusted to approximately 13 with 6N NaOH solution andextracted with diethyl ether (2×150 mL). The combined organic phase waswashed with water, dried over anhydrous Na₂SO₄, and concentrated underreduced pressure to give3-(4-isopropylpiperazin-1-yl)-5-methoxybenzaldehyde (1.24 g, 27%) as aviscous red oil: ¹H NMR (400 MHz, CDCl₃): δ 9.90 (s, 1H), 7.05 (dd,J=2.15, 1.37 Hz, 1H), 6.88 (dd, J=1.95, 1.17 Hz, 1H), 6.69 (t, J=2.34Hz, 1H), 3.85 (s, 3H), 3.25-3.28 (m, 4H), 2.64-2.78 (m, 5H), 1.10 (d,J=6.64 Hz, 6H).

To a solution of 3-(4-isopropylpiperazin-1-yl)-5-methoxybenzaldehyde(1.23 g, 4.68 mmol) in N,N-dimethylacetamide (20 mL) was added2-amino-4,6-dimethoxybenzamide (0.59 g, 3.00 mmol), NaHSO₃ (58.5 wt %,0.87 g, 4.80 mmol) and p-toluenesulfonic acid monohydrate (1.82 g, 9.60mmol). The reaction mixture was stirred at 120° C. for 20 h undernitrogen. After that time the reaction was cooled to rt and concentratedunder reduced pressure. The residue was diluted with saturated Na₂CO₃solution to adjust the pH to 12. The precipitated solids were collectedby filtration, washed with water and dried under vacuum. The product waspurified by flash column chromatography (silica gel, 86:10:4dichloromethane/ethyl acetate/methanol followed by 86:10:4dichloromethane/ethyl acetate/7 N NH₃ in methanol) and prep. HPLC togive2-(3-(4-isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7-dimethoxyquinazolin-4(3H)-one(1.04 g, 50%) as a white solid: mp 212-213° C.; ¹H NMR (400 MHz,DMSO-d₆): δ 11.99 (s, 1H), 7.36 (s, 1H), 7.20 (s, 1H), 6.76 (d, J=2.34Hz, 1H), 6.60 (t, J=1.95 Hz, 1H), 6.53 (d, J=1.95 Hz, 1H), 3.90 (s, 3H),3.85 (s, 3H), 3.81 (s, 3H), 3.22 (t, J=4.29 Hz, 4H), 2.68 (quin, J=6.54Hz, 1H), 2.58 (t, J=4.68 Hz, 4H), 1.01 (d, J=6.63 Hz, 6H); ESI MS m/z437 [M−H]⁻.

Example 35,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1yl)pyridin-2-yl)quinazolin-4(3H)-one

To a solution of 2-amino-4,6-dimethoxybenzamide (0.737 g, 3.76 mmol) and6-fluoropicolinaldehyde (0.470 g, 3.76 mmol) in N,N-dimethylacetamide(20 mL), NaHSO₃ (58.5% SO₂ content, 1.0 g, 5.63 mmol) andp-toluenesulfonic acid monohydrate (0.143 g, 0.75 mmol) were added. Thereaction was heated at 120° C. for 20 h. After that time the reactionwas cooled to rt, concentrated under reduced pressure, diluted withwater (50 mL) and saturated NaHCO₃ solution was added to adjust the pHto 8. The precipitated solids were collected by filtration, washed withwater and dried. The product was purified by flash column chromatography(silica gel, 99:1 dichloromethane/methanol to 97:3dichloromethane/methanol) to give2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.550 g,49%) as a yellow solid: ¹H NMR (400 MHz, DMSO-d₆) δ 11.34 (s, 1H),8.28-8.33 (m, 1H), 8.19-8.27 (m, 1H), 7.47 (dd, J=8.01, 1.76 Hz, 1H),6.82 (d, J=2.34 Hz, 1H), 6.62 (d, J=2.34 Hz, 1H), 3.91 (s, 3H), 3.86 (s,3H); ¹⁹F NMR (376 MHz, DMSO-d₆) δ −68.08 (d, J=8.03 Hz); ESI MS m/z 302[M+H]⁺.

A mixture of 2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.100 g, 0.33 mmol), 1-(2-methoxyethyl)piperazine (0.072 g, 0.49 mmol)and 1,1,3,3-tetramethylguanidine (0.096 g, 0.83 mmol) in dry DMSO (2 mL)was heated at 80° C. for 17 h. After that time the reaction mixture wascooled to rt and diluted with ethyl acetate (30 mL). The organic phasewas washed with water and brine, dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The product was purified by flashcolumn chromatography (silica gel, 97:3 dichloromethane/methanol to 95:5dichloromethane/methanol) to give5,7-dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one(0.070 g, 49%) as a yellow solid: mp 99-101° C.; ¹H NMR (400 MHz, CDCl₃)δ 10.32 (s, 1H), 7.85 (d, J=7.42 Hz, 1H), 7.61-7.71 (m, 1H), 6.77-6.87(m, 2H), 6.48 (d, J=1.95 Hz, 1H), 3.99 (s, 3H), 3.94 (s, 3H), 3.64-3.71(m, 4H), 3.59 (t, J=5.47 Hz, 2H), 3.40 (s, 3H), 2.61-2.71 (m, 6H); ESIMS m/z 426 [M+H]⁺.

Example 42-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one

To a solution of 2-amino-4,6-dimethoxybenzamide (0.40 g, 2.04 mmol) and6-bromopyridine-2-carbaldehyde (0.379 g, 2.04 mmol) inN,N-dimethylacetamide (24 mL) was added NaHSO₃ (0.544 g, 3.06 mmol) andp-toluenesulfonic acid monohydrate (0.078 g, 0.408 mmol) at rt. Thereaction mixture was heated at 120° C. for 6 h. After that time thereaction was cooled to rt, concentrated under reduced pressure anddiluted with water (10 mL). The precipitated solids were collected byfiltration, washed with water and dried. The product was purified byflash column chromatography (silica gel, 95:5 dichloromethane/methanol)to give 2-(6-bromopyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.354g, 48%) as a light yellow solid: ¹H NMR (400 MHz, DMSO-d) δ 11.25 (br s,1H), 8.36 (d, J=7.81 Hz, 1H), 7.87-8.04 (m, 2H), 6.81 (s, 1H), 6.62 (s,1H), 3.91 (s, 3H), 3.87 (s, 3H); ESI MS m/z 362 [M+H]⁺.

To a suspension of2-(6-bromopyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.193 g,0.533 mmol) in dioxane (3 mL) was added palladium(II)acetate (0.012 g,0.053 mmol), rac-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (0.0332 g,0.053 mmol) and cesium carbonate (0.261 g, 0.800 mmol) under nitrogen.The reaction mixture was refluxed under nitrogen for 6.5 h. After thattime the reaction was cooled to rt, filtered and concentrated underreduced pressure. The residue was purified by flash columnchromatography (silica gel, 95:5 dichloromethane/methanol and traceamount of ammonium hydroxide) followed by trituration with diethyl etherto give2-(6-(4-(2-hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.043 g, 20%) as a light yellow solid: mp 175-177° C.; ¹H NMR (400 MHz,DMSO-d) δ 11.05 (br s, 1H), 7.65-7.79 (m, 2H), 7.06 (d, J=8.20 Hz, 1H),6.78 (s, 1H), 6.58 (s, 1H), 4.48 (br s, 1H), 3.91 (s, 3H), 3.86 (s, 3H),3.64 (br s, 4H), 3.55 (br s, 2H), 2.54 (br s, 4H), 2.45 (t, 2H); ESI MSm/z 412 [M+H]⁺.

Example 52-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one

To a suspension of2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.108 g,0.36 mmol) in N,N-dimethylformamide (1.5 mL) was added K₂CO₃ (0.15 g,1.07 mmol) and 1-isobutyl-piperazine (88.9 μL, 0.54 mmol). The resultingmixture was heated at 110° C. for 6 h. After that timeN,N-dimethylacetamide (1.5 mL) was added and heating was continued for20 h. After that time the reaction mixture was cooled to rt, dilutedwith ethyl acetate and filtered. The filtrate was concentrated underreduced pressure and the residue was washed with water, and dried underhigh vacuum. The product was purified by flash column chromatography(silica gel, 99:1 dichloromethane/methanol) to give2-(6-(4-isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.052 g, 34%) as a yellow solid: mp 188-189° C.; ¹H NMR (400 MHz,CDCl₃) δ 10.33 (br s, 1H), 7.83 (d, J=7.42 Hz, 1H), 7.65 (dd, J=8.59,7.42, 1H), 6.76-6.88 (m, 2H), 6.48 (d, J=2.34 Hz, 1H), 3.99 (s, 3H),3.94 (s, 3H), 3.64 (t, J=4.88 Hz, 4H), 2.55 (t, J=4.88 Hz, 4H), 2.17 (d,J=7.42 Hz, 2H), 1.75-1.95 (m, 1H), 0.95 (d, J=6.64 Hz, 6H); ESI MS m/z424 [M+H]⁺.

Example 65,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-yl)quinazazolin-4(3H)-one

To a solution of2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.200 g,0.664 mmol) in DMSO (1.5 mL) under nitrogen was added1-[2-(methylsulfonyl)ethyl]piperazine hydrochloride (0.265 g, 0.996mmol) and 1,1,3,3-tetramethylguanidine. The reaction was stirred at 80°C. for 18 h. After that time the reaction was cooled to rt, diluted withethyl acetate (50 mL), washed with water (2×20 mL), brine, dried overNa₂SO₄ and concentrated under reduced pressure. The product was purifiedby flash column chromatography (silica gel, 95:5dichloromethane/methanol) to give5,7-dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one(0.120 g, 38%) as a yellow solid: mp 243-244° C.; ¹H NMR (400 MHz,CDCl₃): δ 10.28 (br s, 1H), 7.88 (d, J=7.3 Hz, 1H), 7.73-7.66 (m, 1H),6.87-6.81 (m, 2H), 6.49 (d, J=2.2 Hz, 1H), 3.99 (s, 3H), 3.94 (s, 3H),3.69-3.61 (m, 4H), 3.23 (t, J=6.4 Hz, 2H), 3.09 (s, 3H), 2.98 (t, J=6.4Hz, 2H), 2.72-2.64 (m, 4H).

Example 7 Methyl2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetate

To a stirred suspension of methyl 2-(piperazin-1-yl)acetatedihydrochloride (0.67 g, 2.92 mmol) in dry DMSO (1.5 mL) was added1,1,3,3-tetramethylguanidine (0.15 g, 1.07 mmol). The resulting mixturewas stirred at rt for 10 min. Then2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.40 g, 1.33mmol) was added as a solid. The reaction was heated at 90° C. for 4.5 h.After that time the reaction was cooled to rt and diluted with ethylacetate (30 mL). The organic phase was washed with water and brine,dried over sodium sulfate, filtered and concentrated under reducedpressure. The product was purified by flash column chromatography(silica gel, 98:2 dichloromethane/methanol) followed by trituration withdiethyl ether to give methyl2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetate(0.357 g, 61%) as a light yellow solid: mp 176-177° C.; ¹H NMR (400 MHz,CDCl₃) δ 10.30 (br s, 1H), 7.86 (d, J=7.03 Hz, 1H), 7.63-7.71 (m, 1H),6.80-6.86 (m, 2H), 6.48 (d, J=2.34 Hz, 1H), 3.99 (s, 3H), 3.94 (s, 3H),3.77 (s, 3H), 3.67-3.73 (m, 4H), 3.34 (s, 2H), 2.73-2.79 (m, 4H); ESI MSm/z 440 [M+H]⁺.

Example 82-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one

A mixture of 2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.202 g, 0.67 mmol), 2-(piperazin-1-yl)propan-1-ol dihydrochloride(0.292 g, 1.34 mmol) and 1,1,3,3-tetramethylguanidine (0.310 g, 2.69mmol) in dry DMSO (1.5 mL) was heated at 85° C. for 6 h. After that timethe reaction was cooled to rt, diluted with water (15 mL) and extractedwith ethyl acetate (2×30 mL). The combined organic phase was washed withbrine, dried over sodium sulfate, filtered and concentrated underreduced pressure. The product was purified by flash columnchromatography (silica gel, 97:3 dichloromethane/methanol to 95:5dichloromethane/methanol) to give2-(6-(4-(1-hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.155 g, 54%) as a yellow solid: mp 202-204° C.; ¹H NMR (400 MHz,CDCl₃) δ 10.30 (s, 1H), 7.87 (d, J=7.42 Hz, 1H), 7.64-7.72 (m, 1H),6.79-6.88 (m, 2H) 6.49 (d, J=2.34 Hz, 1H) 3.99 (s, 3H), 3.94 (s, 3H),3.61-3.72 (m, 4H), 3.38-3.52 (m, 2H), 2.89-2.99 (m, 1H), 2.78-2.88 (m,2H), 2.54-2.64 (m, 2H), 0.97 (d, J=6.64 Hz, 3H); ESI MS m/z 426 [M+H]⁺.

Example 92-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanamide

To a suspension of 2-(piperazin-1-yl)propanamide dihydrochloride (0.305g, 1.33 mmol) in DMSO (1.5 mL), was added 1,1,3,3,-tetramethylguanidine(0.398 g, 3.45 mmol) at rt. The reaction mixture was stirred for 15 min.After that time2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.20 g,0.664 mmol) was added as a solid in a single portion. The resultingmixture was stirred at 90° C. for 3.5 h. After that time the reactionwas cooled to rt and diluted with dichloromethane (50 mL). The organicphase was washed with brine, dried over Na₂SO₄ and concentrated underreduced pressure. The residue was purified by flash columnchromatography (silica gel, 93:7 dichloromethane/methanol) to give2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanamide(0.080 g, 28%) as a light yellow solid: mp 149-150° C.; ¹H NMR (400 MHz,DMSO-d₆) δ 11.05 (br s, 1H), 7.65-7.79 (m, 2H), 7.30 (s, 1H), 7.06 (d,J=8.20 Hz, 1H), 6.78 (s, 1H), 6.58 (s, 1H), 4.48 (br s, 1H), 3.91 (s,3H), 3.86 (s, 3H), 3.64 (br s, 4H), 3.55 (br s, 2H), 2.54 (br s, 4H),2.45 (t, 2H); ESI MS m/z 439 [M+H]⁺.

Example 102-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)aceticacid

To a solution of methyl2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetate(0.156 g, 0.35 mmol) in THF (10 mL), water (5 mL) and methanol (3 mL)was added lithium hydroxide (0.043 g, 1.77 mmol). The reaction wasstirred for 2 h at rt. After that time the reaction was cooled to rt,concentrated under reduced pressure, diluted with water (5 mL) andacidified with 1N HCl to pH 4.5-5.0. The precipitated solids werecollected by filtration, washed with water and dried under high vacuumto give2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)aceticacid (0.135 g, 91%) as a yellow solid: mp 225° C. dec.; ¹H NMR (400 MHz,DMSO-d₆) δ 11.08 (s, 1H), 7.72-7.79 (m, 1H), 7.67-7.72 (m, 1H), 7.07 (d,J=8.59 Hz, 1H), 6.78 (d, J=1.56 Hz 1H), 6.58 (d, J=1.56 Hz, 1H) 3.91 (s,3H), 3.86 (s, 3H), 3.63-3.74 (m, 4H), 3.23 (s, 2H), 2.65-2.74 (m, 4H);ESI MS m/z 426 [M+H]⁺.

Example 113-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoicacid

To a stirred suspension of methyl 3-(piperazin-1-yl)propanoatedihydrochloride (0.358 g, 1.46 mmol) and2-(6-fluoropyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (0.220 g,0.73 mmol) in dry DMSO (1.5 mL) was added 1,1,3,3-tetramethylguanidine(0.336 g, 2.92 mmol). The resulting mixture was heated at 85° C. for 5h. After that time the reaction was cooled to rt, diluted with water (20mL) and extracted with ethyl acetate (3×25 mL). The organic phase waswashed with water and brine, dried over sodium sulfate, filtered andconcentrated under reduced pressure. The product was purified by flashcolumn chromatography (silica gel, 99:1 dichloromethane/methanol to 97:3dichloromethane/methanol) to give methyl3-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoate(0.220 g, 66%) as an off-white solid: ¹H NMR (400 MHz, CDCl₃) δ 10.31(s, 1H), 7.85 (d, J=7.42 Hz, 1H), 7.62-7.70 (m, 1H), 6.79-6.85 (m, 2H),6.48 (d, J=2.34 Hz, 1H), 3.99 (s, 3H), 3.94 (s, 3H), 3.72 (s, 3H),3.59-3.67 (m, 4H), 2.75-2.83 (m, 2H), 2.54-2.66 (m, 6H); ESI MS m/z 454[M+H]⁺.

To a solution of methyl3-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoate(0.200 g, 0.44 mmol) in THF (10 mL), water (5 mL) and methanol (3 mL)was added lithium hydroxide (0.053 g, 2.21 mmol). The reaction wasstirred for 2 h at rt. After that time the reaction was concentratedunder reduced pressure, diluted with water (5 mL) and acidified using 1NHCl to pH 4.5-5.0. The precipitated solids were collected by filtration,washed with water and dried under high vacuum to give3-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoicacid (0.160 g, 83%) as a yellow solid: mp 241° C. dec.; ¹H NMR (400 MHz,DMSO-d₆) δ 11.15 (s, 1H), 7.68-7.80 (m, 2H), 7.09 (d, J=8.20 Hz, 1H),6.78 (d, J=1.95 Hz, 1H), 6.58 (d, J=1.95 Hz, 1H), 3.91 (s, 3H), 3.86 (s,3H), 3.73 (br s, 4H), 2.77-2.85 (m, 2H), 2.73 (br s, 4H), 2.55 (t,J=7.22 Hz, 2H); ESI MS m/z 440 [M+H]⁺.

Example 122-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-onebis(trifluoroacetate)

PBr₃ (0.160 mL, 1.70 mmol) was added dropwise to a solution of2-(5-(2-hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.40 g, 0.85 mmol) in N,N-dimethylformamide (10 mL) under nitrogen. Theresulting mixture was heated at 60° C. for 2 h. After that time thereaction was cooled to rt and concentrated under reduced pressure andre-dissolved in chloroform (12 mL). Isopropylamine (4 mL) was added andthe resulting mixture was heated at 50° C. in a sealed tube for 24 h.After that time the reaction was cooled to rt and concentrated underreduced pressure. The product was purified by flash columnchromatography (silica gel, 90:10:0.8 dichloromethane/methanol/ammoniumhydroxide) followed by preparative HPLC (0.1% trifluoroacetic acid inacetonitrile/water) to give2-(5-(2-(isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-onebis(trifluoroacetate) (0.056 g, 9%) as a yellow solid: mp 186-188° C.;¹H NMR (400 MHz, DMSO-d₆) δ 11.07 (br s, 1H), 9.56 (br s, 1H), 8.74 (brs, 2H), 8.01 (d, J=8.8 Hz, 1H), 7.59 (d, J=8.4 Hz, 1H), 6.75 (d, J=2.4Hz, 1H), 6.57 (d, J=2.4 Hz, 2H), 4.48 (d, J=11.6 Hz, 2H), 4.37 (dd,J=4.4, 5.2 Hz, 2H), 3.91 (s, 3H), 3.86 (s, 3H), 360-3.40 (m, 5H),3.23-3.10 (m, 4H), 1.31 (d, J=6.8 Hz, 6H), 1.28 (d, J=6.4 Hz, 6H); ¹⁹FNMR (376 MHz, DMSO-d₆) δ ppm −73.80; ESI MS m/z 511 [M+H]⁺.

Example 132-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneacetate

To a mixture of(6-bromo-5-(2-((tert-butyldimethylsilyl)oxy)ethoxy)pyridin-2-yl)methanol(2.18 g, 6.0 mmol), 1-isopropylpiperazine (0.92 g, 7.2 mmol), BINAP(0.37 g, 0.6 mmol), and Cs₂CO₃ (3.18 g, 9.0 mmol) in nitrogen saturatedtoluene (40 mL) was added palladium(II) acetate (0.13 g, 0.6 mmol). Theresulting mixture was heated at 100° C. for 18 h. After that time thereaction was cooled to rt, diluted with brine and extracted with ethylacetate. The organic phase was dried over Na₂SO₄ and concentrated underreduced pressure. The product was purified by flash columnchromatography (silica gel, 95:5 dichloromethane/methanol to 90:10dichloromethane/methanol) to give(5-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)methanol(1.07 g, 44%) as a sticky yellow solid: ¹H NMR (400 MHz, CDCl₃): δ 7.05(d, J=7.6 Hz, 1H), 6.66 (d, J=8.0 Hz, 1H), 4.59 (s, 2H), 4.06-3.97 (m,4H), 3.53 (br s, 4H), 2.68 (m, 5H), 1.10 (d, J=6.4 Hz, 6H), 0.90 (s,9H), 0.09 (s, 6H).

To a solution of(5-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)methanol(1.07 g, 2.61 mmol) in 1,2-dichloroethane (20 mL) under nitrogen wasadded 2-iodoxybenzoic acid (0.88 g, 3.14 mmol) at rt. The reactionmixture was stirred at 80° C. for 1.5 h. After that time the reactionwas cooled to rt and concentrated under reduced pressure. The productwas purified by flash column chromatography (silica gel, 95:5dichloromethane/methanol) to give5-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-6-(4-isopropylpiperazin-1-yl)picolinaldehyde(0.63 g, 59%) as a brown solid: ¹H NMR (400 MHz, CDCl₃): δ 9.85 (s, 1H),7.59 (d, J=8.4 Hz, 1H), 7.17 (d, J=8.0 Hz, 1H), 4.19-4.15 (m, 2H),4.02-3.88 (m, 6H), 3.56-3.48 (m, 1H), 3.20 (br s, 4H), 1.37 (d, J=6.4Hz, 6H), 0.90 (s, 9H), 0.09 (s, 6H); ESI MS m/z 408 [M+H]⁺.

To a solution of 2-amino-4,6-dimethoxy-benzamide (0.302 g, 1.54 mmol) inN,N-dimethylacetamide (10 mL) was added5-[2-(tert-butyl-dimethyl-silanyloxy)-ethoxy]-6-(4-methanesulfonyl-2-trifluoromethyl-phenyl)-pyridine-2-carbaldehyde(0.630 g, 1.54 mmol) followed by NaHSO₃ (0.321 g, 3.08 mmol) andp-toluenesulfonic acid monohydrate (0.293 g, 1.54 mmol). The resultingmixture was heated at 120° C. for 20 h. After that time the reaction wascooled to rt, concentrated under reduced pressure and diluted withwater. Saturated Na₂CO₃ was added to adjust the pH to 9-10. Theprecipitated solids were collected by filtration, washed with water anddried under vacuum. The product was purified by flash columnchromatography (silica gel, 90:10:1 dichloromethane/methanol/ammoniumhydroxide) to give2-(5-(2-hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.47 g, 65%) as a sticky yellow solid: ¹H NMR (400 MHz, CDCl₃): δ 10.22(br s, 1H), 8.07 (d, J=8.0 Hz, 1H), 7.23 (d, J=8.4 Hz, 1H), 6.78 (d,J=2.4 Hz, 1H), 6.45 (d, J=2.4 Hz, 1H), 4.21 (dd, J=4.0, 4.4 Hz, 2H),3.98 (s, 3H), 3.93 (s, 3H), 3.55-3.53 (m, 5H), 2.76-2.72 (m, 6H), 1.11(d, J=6.8 Hz, 6H); ESI MS m/z 470 [M+H]⁺.

A sample of2-(5-(2-hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.050 g,) was further purified by preparative HPLC (0.1% formic acid inacetonitrile/water) to give2-(5-(2-hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneformate (0.026 g, 52%) as a yellow solid: mp 138-139° C.; ¹H NMR (400MHz, DMSO-d₆): δ 10.62 (br s, 1H), 8.29 (s, 1H), 7.89 (d, J=8.4 Hz, 1H),7.43 (d, J=8.8 Hz, 1H), 6.75 (d, J=2.4 Hz, 1H), 6.54 (d, J=2.4 Hz, 1H),4.12 (t, J=4.8 Hz, 2H), 3.90 (s, 3H), 3.85 (s, 3H), 3.78 (t, J=4.8 Hz,2H), 3.56-3.30 (m, 8H), 2.72-2.66 (m, 1H), 2.62-2.58 (m, 2H), 1.02 (d,J=6.8 Hz, 6H); ESI MS m/z 470 [M+H]⁺.

Example 142-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one

To a solution of 2-bromo-6-(hydroxymethyl)pyridin-3-ol (2.04 g, 10 mmol)and K₂CO₃ (2.07 g, 15 mmol) in acetone (50 mL) was added iodomethane(1.84 g, 13 mmol) at rt. The reaction mixture was refluxed for 4 h.After that time the reaction was cooled to rt, filtered to remove solidsand concentrated under reduced pressure to give(6-bromo-5-methoxypyridin-2-yl)methanol (3.42 g, >99%) as a white solid:¹H NMR (400 MHz, CDCl₃): δ 7.25 (d, J=8.0 Hz, 1H), 7.16 (d, J=8.4 Hz,1H), 4.70 (s, 2H), 3.93 (s, 3H), 2.78 (br s, 1H).

To a solution of (6-bromo-5-methoxypyridin-2-yl)methanol (2.30 g, 10mmol) in dichloromethane (50 mL) was added MnO₂ (8.0 g) at rt. Thereaction mixture was stirred at rt for 24 h. After that time thereaction was cooled to rt, filtered to remove solids and concentratedunder reduced pressure to give 6-bromo-5-methoxypicolinaldehyde (1.35 g,63%) as an off-white solid: ¹H NMR (400 MHz, CDCl₃): δ 9.94 (s, 1H),7.96 (d, J=8.4 Hz, 1H), 7.27 (d, J=8.0 Hz, 1H), 4.03 (s, 3H).

To a solution of 2-amino-4,6-dimethoxy-benzamide (1.23 g, 6.25 mmol) inN,N-dimethylacetamide (100 mL) was added6-bromo-5-methoxypicolinaldehyde (1.35 g, 1.2 mmol) followed by NaHSO₃(0.98 g, 9.38 mmol) and p-toluenesulfonic acid monohydrate (1.19 g, 6.25mmol). The resulting mixture was heated at 120° C. for 20 h. After thattime the reaction was cooled to it, concentrated under reduced pressureand diluted with saturated Na₂CO₃. The precipitated solids werecollected by filtration, washed with water and dried under vacuum. Theproduct was purified by flash column chromatography (silica gel, 99:1dichloromethane/methanol) followed by trituration with ethyl acetate togive 2-(6-bromo-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(1.27 g, 52%) as a white solid: ¹H NMR (400 MHz, DMSO-d₆): δ 11.03 (brs, 1H), 8.32 (d, J=8.4 Hz, 1H), 7.72 (d, J=8.8 Hz, 1H), 6.76 (d, J=2.0Hz, 1H), 6.56 (d, J=2.4 Hz, 1H), 4.00 (s, 3H), 3.90 (s, 3H), 3.85 (s,3H).

To a mixture of2-(6-bromo-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.196 g, 0.50 mmol), 1-isopropylpiperazine (0.077 g, 0.60 mmol), BINAP(0.062 g, 0.10 mmol), and Cs₂CO₃ (0.652 g, 2.0 mmol) in nitrogensaturated dioxane (10 mL) was added palladium(II) acetate (0.022 g, 0.10mmol). The resulting mixture was heated at 100° C. for 3 h. After thattime the reaction was cooled to rt, diluted with dichloromethane (100mL) and washed with water (20 mL). The organic phase was dried overNa₂SO₄ and concentrated under reduced pressure. The product was purifiedby flash column chromatography (silica gel, 90:10:0.1dichloromethane/methanol/ammonium hydroxide) to give2-(6-(4-isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(0.055 g, 25%) as a pale yellow solid: mp 208-210° C.; ¹H NMR (400 MHz,CDCl₃): δ 10.23 (br s, 1H), 8.08 (d, J=8.0 Hz, 1H), 7.17 (d, J=8.0 Hz,1H), 6.79 (d, J=2.4 Hz, 1H), 6.45 (d, J=2.4 Hz, 1H), 3.98 (s, 3H), 3.95(s, 3H), 3.93 (s, 3H), 3.56 (br s, 4H), 2.78 (br s, 5H), 1.15 (d, J=5.6Hz, 6H); ESI MS m/z 440 [M+H]⁺.

Example 155,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-oneHydrochloride

Preparation of 6-(4-Methylpiperazin-1-yl)picolinaldehyde (3). Asuspension of 6-bromopicolinaldehyde (1.50 g, 8.10 mmol),1-methylpiperazine (2, 4.04 g, 40.0 mmol), potassium carbonate (4.46 g,32.0 mmol) and anhydrous CH₃CN (10 mL) was placed in a sealed vessel andthe mixture was heated at 110° C. for 17 h. The mixture was diluted withwater (50 mL) and brought to pH 7 using 2 N aq. HCl (15 mL). Theresulting solution was extracted with CH₂Cl₂ (3×50 mL) and the combinedextracts were washed with brine (200 mL). The solution was dried(Na₂SO₄), filtered, concentrated and purified by silica gelchromatography eluting with 0-5% CH₃OH in CH₂Cl₂ to afford the titlecompound (0.528 g, 32%) as an orange oil: ¹H NMR (500 MHz, CDCl₃) δ 9.84(s, 1H), 7.62 (t, J=8.0 Hz, 1H), 7.27 (d, J=7.0 Hz, 1H), 6.85 (d, J=7.0Hz, 1H), 3.67-3.65 (m, 4H), 2.55-2.52 (m, 4H), 2.36 (s, 3H).

Preparation of5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-oneHydrochloride (Example 15). 2-Amino-4,6-dimethoxybenzamide (4, 200 mg,1.02 mmol), 6-(4-methylpiperazin-1-yl)picolinaldehyde (3, 231 mg, 1.12mmol), para-toluenesulfonic acid (466 mg, 2.45 mmol), sodium hydrogensulfite (297 mg, 2.85 mmol) and anhydrous N,N-dimethylacetamide (12 mL)were mixed in a sealed tube and heated at 110° C. for 17 h. The solventwas removed in vacuo, the residue was dissolved in methanol (20 mL) andDCM:CH₃OH:aq. NH₄OH (80:18:2, 20 mL) and the contents were adsorbed ontosilica gel (5 g). The adsorbed material was purified by silica gelchromatography eluting with 0-50% DCM:CH₃OH:aq. NH₄OH (80:18:2) inCH₂Cl₂ then using preparative HPLC. After concentration, 5 N aq. HCl (3mL) was added and the suspension was concentrated in vacuo and placed ina drying oven at 50° C. for 17 h to afford the title compound (0.127 g,25%) as an off-white solid: ¹H NMR (500 MHz, CD₃OD) δ 7.97-7.95 (m, 1H),7.82 (d, J=7.5 Hz, 1H), 7.39 (d, J=8.5 Hz, 1H), 7.20 (d, J=2.0 Hz, 1H),6.79 (d, J=2.0 Hz, 1H), 4.82 (d, J=14.5 Hz, 2H), 4.00 (s, 3H), 3.99 (s,3H), 3.67 (d, J=12.5 Hz, 2H), 3.48-3.42 (m, 2H), 3.31-3.24 (m, 2H), 3.00(s, 3H); ESI MS m/z 382 [M+H]⁺.

Example 16 Preparation of5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-oneHydrochloride

Preparation of 6-Morpholinopicolinaldehyde (6): Following the methoddescribed for 3 above (see Example 15), compound 6 was made frommorpholine (5) in 34% yield: ¹H NMR (500 MHz, CDCl₃) δ 9.89 (s, 1H),7.67-7.65 (m, 1H), 7.31 (d, J=7.0 Hz, 1H), 6.84 (d, J=8.5 Hz, 1H),3.86-3.84 (m, 4H), 3.62-3.60 (m, 4H).

Preparation of5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-oneHydrochloride (Example 16): Following the method described for Example15 above, Example 16 was made from 6-morpholinopicolinaldehyde (6) in21% yield: ¹H NMR (500 MHz, DMSO-d₆) δ 11.0 (br s, 1H), 7.77 (t, J=7.5Hz, 1H), 7.72 (d, J=7.0 Hz, 1H), 7.06 (d, J=8.5 Hz, 1H), 6.77 (d, J=2.0Hz, 1H), 6.57 (d, J=2.0 Hz, 1H), 3.90 (s, 3H), 3.86 (s, 3H), 3.75-3.73(m, 4H), 3.63-3.61 (m, 4H); ESI MS m/z 369 [M+H]⁺.

Example 17 Preparation of2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride

Preparation of 6-(4-isopropylpiperazin-1-yl)picolinaldehyde (8):Following the method described for 3 above (see Example 15), compound 8was made from 1-isopropylpiperazine (7) in 22% yield: ¹H NMR (500 MHz,CDCl₃) δ 9.89 (s, 1H), 7.60 (t, J=7.5 Hz, 1H), 7.27 (d, J=7.0 Hz, 1H),6.84 (d, J=8.5 Hz, 1H), 3.66-3.64 (m, 4H), 2.74 (sept, 1H), 2.66-2.64(m, 4H).

Preparation of2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride (Example 17): Following the method described for Example15 above, compound Example 17 was made from6-(4-isopropylpiperazin-1-yl)picolinaldehyde (8) in 23% yield: ¹H NMR(500 MHz, DMSO-d₆) δ 10.7 (br s, 1H), 7.83 (t, J=7.5 Hz, 1H), 7.78 (d,J=7.5 Hz, 1H), 7.17 (d, J=8.0 Hz, 1H), 6.79 (d, J=2.0 Hz, 1H), 6.59 (d,J=2.0 Hz, 1H), 4.76-4.72 (m, 2H), 3.90 (s, 3H), 3.87 (s, 3H), 3.53-3.51(m, 3H), 3.41-3.36 (m, 2H), 3.10-3.06 (m, 2H), 1.33 (d, J=7.0 Hz, 6H);ESI MS m/z 410 [M+H]⁺.

Example 18 Preparation of5,7-Dimethoxy-2-(6-(4-(methylsulfonyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-oneHydrochloride

Preparation of 6-(4-(Methylsulfonyl)piperazin-1-yl)picolinaldehyde (10):A suspension of 6-bromopicolinaldehyde (1.50 g, 8.10 mmol),1-(methylsulfonyl)piperazine (9, 5.30 g, 32.2 mmol), potassium carbonate(4.45 g, 32.2 mmol) and anhydrous DMF (10 mL) was placed in a sealedvessel and the mixture was heated at 120° C. for 20 h. The mixture wasdiluted with water (50 mL) and brought to pH 7 using 2 N aq. HCl (15mL). The resulting solution was extracted with CH₂Cl₂ (3×50 mL) and thecombined extracts were washed with brine (200 mL). The solution wasdried (MgSO₄), filtered, concentrated, and purified by silica gelchromatography eluting with 0-3% CH₃OH in CH₂Cl₂ to afford the titlecompound (0.441 g, 20%) as an orange oil: ¹H NMR (500 MHz, CDCl₃) δ 9.90(s, 1H), 7.67 (t, J=8.0 Hz, 1H), 7.34 (d, J=7.0 Hz, 1H), 6.89 (d, J=8.5Hz, 1H), 3.80-3.78 (m, 4H), 3.37-3.33 (m, 4H), 2.81 (s, 3H).

Preparation of5,7-Dimethoxy-2-(6-(4-(methylsulfonyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-oneHydrochloride (Example 18): Following the method described for Example15 above, Example 18 was made from6-(4-(methylsulfonyl)piperazin-1-yl)picolinaldehyde (10) in 11% yield:¹H NMR (500 MHz, DMSO-d₆) δ 7.81-7.78 (m, 1H), 7.75-7.54 (m, 1H), 7.14(d, J=8.0 Hz, 1H), 6.83 (d, J=2.0 Hz, 1H), 6.59 (d, J=2.0 Hz, 1H), 3.91(s, 3H), 3.87 (s, 3H), 3.83-3.81 (m, 4H), 3.24-3.22 (m, 4H), 2.92 (s,3H); ESI MS m/z 446 [M+H]⁺.

Example 19 Preparation of5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-onedihydrochloride

Preparation of Benzyl 4-propionylpiperazine-1-carboxylate (17): Asolution of benzyl piperazine-1-carboxylate (16, 10.0 g, 45.4 mmol),triethylamine (6.89 g, 68.0 mmol) and dry CH₂Cl₂ (150 mL) was cooled to0° C. and propionyl chloride (4.62 g, 50.0 mmol) was added dropwise over10 min. The reaction was kept under nitrogen and allowed to warm to roomtemperature. After stirring at rt for 17 h, water (50 mL) was added andthe aqueous layer was extracted with CH₂Cl₂ (2×50 mL). The solution wasdried over Na₂SO₄, filtered and concentrated to a yellow oil. Theproduct was purified by silica gel chromatography eluting with 0-3%CH₃OH in CH₂Cl₂ to afford the title compound (9.08 g, 72%) as a clear,colorless oil: ¹H NMR (500 MHz, CDCl₃) δ 7.40-7.30 (m, 5H), 5.15 (s,2H), 3.70-3.35 (m, 8H), 2.35 (q, J=7.5 Hz, 2H), 1.15 (t, J=7.5 Hz, 3H).

Preparation of 1-(Piperazin-1-yl)propan-1-one (18): A Parr shaker bottlewas charged with a suspension of benzyl4-propionylpiperazine-1-carboxylate (16), 10% palladium on carbon, 50%wet (900 mg) and absolute ethanol (100 mL). The Parr bottle was filledwith 20 psi of hydrogen and shook for 3 h at rt. The resultingsuspension was filtered through Celite and concentrated to yield thetitle compound (4.31 g, 93%) as a clear, colorless oil: ¹H NMR (500 MHz,CDCl₃) δ 3.75-3.63 (m, 2H), 3.48-3.42 (m, 2H), 2.87-2.80 (m, 4H), 2.34(q, J=7.5 Hz, 2H), 1.98 (br s, 1H), 1.15 (t, J=7.5 Hz, 3H).

Preparation of 6-(4-Propionylpiperazin-1-yl)picolinaldehyde (19): To ahigh-pressure tube was added 6-bromopicolinaldehyde (1, 2.00 g, 10.7mmol), 1-(piperazin-1-yl)propan-1-one (18, 3.82 g, 26.9 mmol), potassiumcarbonate (6.69 g, 48.4 mmol) and anhydrous DMF (12 mL). The tube wassealed and the reaction mixture was heated at 120° C. for 17 h. Thereaction suspension was poured into water (100 mL) and the mixture wasbrought to pH 7 using 2 N aq. HCl (20 mL). The solution was extractedwith CH₂Cl₂ (3×100 mL), washed with brine (300 mL), dried over Na₂SO₄,filtered and concentrated to a brown oil. The product was purified bysilica gel chromatography eluting with 0-5% CH₃OH in CH₂Cl₂ to affordthe title compound (0.815 g, 31%) as an orange solid: ¹H NMR (500 MHz,CDCl₃) δ 9.89 (s, 1H), 7.68 (t, J=8.0 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H),6.84 (d, J=8.0 Hz, 1H), 3.84-3.73 (m, 2H), 3.68-3.59 (m, 2H), 2.40 (q,J=7.5 Hz, 2H), 1.19 (t, J=7.5 Hz, 3H).

Preparation of5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-onedihydrochloride (Example 19): Following the method described for Example15 above, Example 19 was made from6-(4-propionylpiperazin-1-yl)picolinaldehyde (19) in 15% yield: ¹H NMR(500 MHz, CD₃OD) δ 7.91 (t, J=8.0 Hz, 1H), 7.70 (d, J=7.5 Hz, 1H), 7.31(d, J=8.5 Hz, 1H), 7.10 (d, J=2.0 Hz, 1H), 6.83 (d, J=2.0 Hz, 1H), 4.02(s, 3H), 4.01 (s, 3H), 3.90-3.72 (m, 8H), 2.51 (q, J=7.5 Hz, 2H), 1.17(t, J=7.5 Hz, 3H); ESI MS m/z 424 [M+H]⁺.

Example 20 Preparation of2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one

Preparation of 2-(3-Bromo-5-(trifluoromethoxy)phenyl)-1,3-dioxolane(21): A solution of 3-bromo-5-trifluoromethoxybenzaldehyde (20, 2.00 g,7.43 mmol), ethylene glycol (2.31 g, 37.2 mmol), para-toluenesulfonicacid (50 mg, 0.26 mmol) in anhydrous toluene (50 mL) was refluxed usinga Dean-Stark apparatus for 3 h. The solution was concentrated and theproduct purified by silica gel chromatography eluting with 0-10% EtOAcin hexanes to afford the title compound (1.25 g, 54%) as a clear,colorless oil: ¹H NMR (500 MHz, CDCl₃) δ 7.57 (t, J=1.5 Hz, 1H), 7.37(s, 1H), 7.28 (s, 1H), 5.79 (s, 1H), 4.12-4.01 (m, 4H).

Preparation of1-(3-(1,3-Dioxolan-2-yl)-5-(trifluoromethoxy)phenyl)-4-isopropylpiperazine(22): Tris(dibenzylideneacetone)dipalladium(0) (363 mg, 0.396 mmol) wasadded to a suspension of2-(3-bromo-5-(trifluoromethoxy)phenyl)-1,3-dioxolane (21, 1.24 g, 3.96mmol), isopropylpiperazine (610 mg, 4.75 mmol),racemic-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (rac-BINAP, 345 mg,0.554 mmol), cesium carbonate (2.58 g, 7.92 mmol) and anhydrous toluene(50 mL). The mixture was placed under nitrogen and heated to 100° C. for17 h. The solvent was removed in vacuo and methanol (30 mL) and silicagel (30 g) were added. After removing the methanol the adsorbed crudeproduct was purified by silica gel chromatography eluting with 0-60%EtOAc in hexanes to afford the title compound (996 mg, 70%) as an orangeoil: ¹H NMR (500 MHz, CDCl₃) δ 6.93 (s, 1H), 6.78 (s, 1H), 6.68 (s, 1H),5.77 (s, 1H), 4.17-3.99 (m, 4H), 3.26-3.22 (m, 4H), 2.72 (sept, 1H),2.68-2.62 (m, 4H), 1.08 (d, J=6.5 Hz, 6H).

Preparation of3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)benzaldehyde (23):Water (1.3 mL) was added to a suspension of1-(3-(1,3-dioxolan-2-yl)-5-(trifluoromethoxy)phenyl)-4-isopropylpiperazine(22, 996 mg, 2.76 mmol) and formic acid (6.71 g, 145 mmol). The mixturewas heated to 60° C. for 20 h. The reaction mixture was concentrated toa brown oil in vacuo. The oil was dissolved in EtOAc (100 mL) and thesolution was washed with saturated aq. NaHCO₃ solution (100 mL) andbrine (100 mL). After drying over Na₂SO₄, the suspension was filteredand the filtrate concentrated to an orange oil. The residue was purifiedby silica gel chromatography eluting with 0-80% EtOAc in hexanes toafford the title compound (608 mg, 70%) as an orange oil: ¹H NMR (500MHz, CDCl₃) δ 9.92 (s, 1H), 6.93 (s, 1H), 7.31 (s, 1H), 7.13 (s, 1H),6.92 (s, 1H), 3.30-3.28 (m, 4H), 2.73 (sept, 1H), 2.70-2.64 (m, 4H),1.10 (d, J=6.5 Hz, 6H).

Preparation of2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one(Example 20): Following the method described for Example 15 above,Example 20 was made from3-(4-isopropylpiperazin-1-yl)-5-(trifluoromethoxy)benzaldehyde (23) in49% yield: ¹H NMR (500 MHz, DMSO-d₆) δ 12.1 (s, 1H), 7.72 (s, 1H), 7.51(s, 1H), 7.01 (s, 1H), 6.77 (d, J=2.0 Hz, 1H), 6.55 (d, J=2.0 Hz, 1H),3.90 (s, 3H), 3.87 (s, 3H), 3.35-3.28 (m, 4H), 2.70 (sept, 1H),2.61-2.56 (m, 4H), 1.02 (d, J=6.5 Hz, 6H). ESI MS m/z 493 [M+H]⁺.

Example 21 Preparation of7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride

Preparation of5,7-Difluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one(8): A mixture of 2-amino-4,6-difluorobenzamide (7, 330 mg, 1.70 mmol),6-(4-isopropylpiperazin-1-yl)picolinaldehyde (8, Scheme 6, 330 mg, 1.42mmol), p-toluenesulfonic acid (590 mg, 3.12 mmol), NaHSO₃ (370 mg, 3.55mmol) and DMA (15 mL) was heated for 24 h at 110° C. under nitrogen. Themixture was partitioned between water (25 mL) and CH₂Cl₂ (25 mL) andshaken vigorously. The layers were separated and the organic phase wasdried over sodium sulfate and filtered. The solvent was removed underreduced pressure and the residue was purified by silica gelchromatography eluting with 0-5% MeOH in CH₂Cl₂ to provide the titlecompound (112 mg, 20%): ¹H NMR (500 MHz, CDCl₃) δ 10.59 (s, 1H), 7.85(d, J=7.3 Hz, 1H), 7.67 (dd, J=7.3, 8.5 Hz, 1H), 7.30-7.26 (m, 1H),6.93-6.89 (m, 1H), 6.86 (d, J=8.5 Hz, 1H), 3.69-3.60 (m, 4H), 2.82-2.73(m, 1H), 2.71-2.66 (m, 4H), 1.12 (s, 3H), 1.11 (s, 3H).

Preparation of7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride (Example 21): To a solution of5,7-difluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one(8, 44 mg, 0.11 mmol) and MeOH (5 mL) was added NaOMe (25% solution inMeOH) at room temperature. After stirring overnight, the solvent wasremoved under reduced pressure and the residue partitioned between waterand CH₂Cl₂. The organic phase was dried over sodium sulfate andfiltered. The solvent was removed under reduced pressure and the residuewas purified by preparative HPLC followed by treatment with aqueous HClto provide the title compound (7.5 mg, 17%) as an white solid: ¹H NMR(500 MHz, CD₃OD) δ 7.90 (d, J=7.2 Hz, 1H), 7.82 (dd, J=7.4, 8.3 Hz, 1H),7.14 (d, J=8.4 Hz, 1H), 7.06-6.99 (m, 1H), 6.93-6.85 (m, 1H), 4.79-4.66(m, 2H), 3.98 (s, 3H), 3.71-3.56 (m, 3H), 3.29-3.19 (m, 3H), 1.44 (s,3H), 1.43 (s, 3H); ESI MS m/z 398 [M+H]⁺.

Example 22 Preparation of7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride

Preparation of7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride (Example 22): To a mixture of NaH (58 mg, 1.44 mmol) andDMSO (2.0 mL) at room temperature was slowly added a solution of benzylalcohol (173 mg, 1.6 mmol) in DMSO (3.0 mL). After stirring the mixturefor 20 min, a solution of7-fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one(9, 65 mg, 0.16 mmol) in DMSO (3 mL) was slowly added. The reactionmixture was then heated at 80° C. for 14 h. The reaction mixture wascooled to room temperature and water (50 mL) was added. The precipitatedsolid was collected by filtration and purified by silica gelchromatography eluting with 0-5% MeOH in CH₂Cl₂ followed by treatmentwith aqueous HCl to provide the title compound (38 mg, 49%) as anoff-white solid: ¹H NMR (500 MHz, CD₃OD) δ 7.95-7.88 (m, 1H), 7.83 (d,J=7.4 Hz, 1H), 7.50-7.46 (m, 2H), 7.43-7.38 (m, 2H), 7.38-7.33 (m, 1H),7.30 (d, J=8.6 Hz, 1H), 7.08 (s, 1H), 6.84 (s, 1H), 5.30 (s, 2H),4.83-4.77 (m, 2H), 3.98 (s, 3H), 3.68-3.57 (m, 3H), 3.42-3.33 (m, 2H),3.29-3.21 (m, 2H), 1.44 (s, 3H), 1.43 (s, 3H); ESI MS m/z 486 [M+H]⁺.

Example 23 Preparation of7-(4-Isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride

Preparation of 2-(6-Bromopyridin-2-yl)-5,7-difluoroquinazolin-4(3H)-one(12): A mixture of 6-bromopicolinaldehyde (2.8 g, 15 mmol),2-amino-4,6-difluorobenzamide (2.5 g, 10 mmol), p-toluenesulfonic acid(4.2 g, 22 mmol), NaHSO₃ (2.6 g, 25 mmol) and DMA (40 mL) was heated at110° C. for 48 h in a sealed tube. The reaction mixture was cooled toroom temperature and partitioned between water and EtOAc (25 mL). Theaqueous phase was extracted with EtOAc (2×25 mL). The organic layerswere combined, dried over sodium sulfate, filtered and the solvent wasremoved under reduced pressure. The product was purified bycrystallization (EtOAc) to provide the title compound (500 mg, 15%): ¹HNMR (500 MHz, DMSO-d₆) δ 11.97 (s, 1H), 8.32 (d, J=7.7, 0.7 Hz, 1H),8.05-7.96 (m, 1H), 7.91 (dd, J=7.7, 0.7 Hz, 1H), 7.46-7.29 (m, 2H).

Preparation of2-(6-bromopyridin-2-yl)-7-fluoro-5-methoxyquinazolin-4(3H)-one (13): Toa mixture of 2-(6-bromopyridin-2-yl)-5,7-difluoroquinazolin-4(3H)-one(12, 1.05 g, 3.11 mmol) and MeOH (15 mL) was added NaOMe (25% solution,50 mL) at room temperature. After 24 hours, another portion of 25% NaOMewas added (25 mL). After an additional 48 hours at room temperature, thereaction was quenched with water and the pH adjusted to 7 with 2N HCl.The volume was reduced by half under reduced pressure and the remainingmixture was extracted with CH₂Cl₂ (3×250 mL). The combined organicphases were dried over sodium sulfate, filtered and concentrated underreduced pressure to provide the title compound (970 mg, 89%) as anoff-white solid. ¹H NMR (500 MHz, CDCl₃) δ 10.56 (s, 1H), 8.57-8.43 (m,1H), 7.79-7.74 (m, 1H), 7.71-7.62 (m, 1H), 7.09-6.99 (m, 1H), 6.71-6.65(m, 1H), 4.02 (s, 3H).

Preparation of7-(4-Isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride (Example 23): A solution of2-(6-bromopyridin-2-yl)-7-fluoro-5-methoxyquinazolin-4(3H)-one (13, 100mg, 0.29 mmol) and 1-isopropylpiperazine (5 mL) was heated at 80° C. for18 hours. Water (25 mL) was added and the mixture was extracted withCH₂Cl₂ (2×25 mL). The combined extracts were dried over sodium sulfateand filtered. The solvent was removed under reduced pressure and theresidue was purified by silica gel chromatography 0-10% MeOH in CH₂Cl₂.The product was further purified by preparative HPLC followed bytreatment with aqueous HCl to provide the title compound (62 mg, 54%) asa white solid: ¹H NMR (300 MHz, CD₃OD) δ 7.85-7.79 (m, 1H), 7.78-7.74(m, 1H), 7.20-7.14 (m, 1H), 6.67-6.59 (m, 1H), 4.78-4.66 (m, 1H),4.24-4.14 (m, 1H), 3.93-3.85 (s, 3H), 3.57-3.36 (m, 16H), 3.19-3.00 (m,4H), 1.33 (s, 6H), 1.32 (s, 6H); ESI MS m/z 506 [M+H]⁺.

Example 24 Preparation of intermediate3-Amino-5-methoxy-[1,1′-biphenyl]-4-carboxamide

Preparation of 4-Bromo-2-fluoro-6-methoxybenzonitrile (2): To a solutionof 4-bromo-2,6-difluorobenzonitrile (1, 15.0 g, 69 mmol) in THF (100 mL)at 0° C. was slowly added sodium methoxide (25% wt in methanol, 14.9 g,69 mmol) and the mixture was stirred at room temperature for 16 h. Themixture was concentrated and partitioned between dichloromethane (500mL) and water (250 mL). The organic extracts were dried (MgSO₄),filtered, and concentrated. The residue was purified by silica gelchromatography eluting with ethyl acetate in hexane (0-100%) to affordthe title compound (15.2 g, 99%) as a white solid: ¹H NMR (500 MHz,CDCl₃) δ 7.01-6.99 (m, 1H), 6.94 (s, 1H), 3.96 (s, 1H).

Preparation of 3-Fluoro-5-methoxy-[1,1′-biphenyl]-4-carbonitrile (3): Amixture of 4-bromo-2-fluoro-6-methoxybenzonitrile (2, 5.22 g, 22.7mmol), phenylboronic acid (4.15 g, 34 mmol) and 2 M Na₂CO₃ (23 mL) intoluene (100 mL) was purged with N₂ for 10 min. Pd(PPh₄)₃ (2.62 g, 27.7mmol) and EtOH (5 drops) were added. The mixture was then heated underN₂ in a sealed tube at 80° C. for 18 h. The mixture was cooled to roomtemperature, diluted with EtOAc (300 mL), filtered through celite,concentrated and purified by silica gel chromatography eluting withethyl acetate in hexane (0-100%) to afford the title compound (5.05 g,97%) as a white solid: ¹H NMR (500 MHz, CDCl₃) δ 7.57-7.55 (m, 2H),7.50-7.23 (m, 3H), 7.01-6.99 (m, 1H), 6.93 (s, 1H), 3.95 (s, 3H).

Preparation of 3-Amino-5-methoxy-[1,1′-biphenyl]-4-carbonitrile (4): Asolution of 3-fluoro-5-methoxy-[1,1′-biphenyl]-4-carbonitrile (3, 4.0 g,17.6 mmol) in ammonia saturated DMSO (100 mL) was heated in an autoclaveat 150° C. for 18 h. After this time, the mixture was cooled to roomtemperature, concentrated and purified by silica gel chromatographyeluting with ethyl acetate in hexane (0-100%) to afford the titlecompound (2.06 g, 52%) as a light yellow solid: ¹H NMR (500 MHz, CDCl₃)δ7.54-7.52 (m, 2H), 7.46-7.43 (m, 2H), 7.41-7.38 (m, 1H), 6.52 (d, J=1.0Hz, 1H), 6.43 (d, J=1.0 Hz, 1H), 4.43 (br s, 2H), 3.93 (s, 3H).

Preparation of 3-Amino-5-methoxy-[1,1′-biphenyl]-4-carboxamide (Example24): A mixture of 3-amino-5-methoxy-[1,1′-biphenyl]-4-carbonitrile (4,0.64 g, 2.86 mmol) and KOH (1.12 g, 20 mmol) in EtOH (24 mL) in a sealedtube was heated at 100° C. for 18 h. Additional KOH (0.56 g, 10 mmol)was added and heating was continued for another 18 h. After this time,the mixture was cooled to room temperature, and the pH was adjusted to^(˜)2 by addition of concentrated HCl. After concentration, the residuewas partitioned between ethyl acetate (100 mL) and water (50 mL). Theextracts were washed with brine (50 mL), dried (Na₂SO₄), filtered, andconcentrated. The residue was purified by silica gel chromatographyeluting with methanol in dichloromethane (0-20%) to afford the titlecompound (0.28 g, 40%) as a yellow solid: ¹H NMR (500 MHz, DMSO-d₆) δ7.62-7.60 (m, 2H), 7.58 (s, 1H), 7.47-7.44 (m, 2H), 7.39-7.36 (m, 1H),7.28 (s, 1H), 6.61 (d, J=1.5 Hz, 1H), 6.56 (s, 2H), 6.43 (d, J=1.5 Hz,1H), 3.87 (s, 3H); ESI MS m/z 243 [M+H]⁺.

Example 25 Preparation of2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxy-7-phenylquinazolin-4(3H)-one

Preparation of 2-Bromo-6-(1,3-dioxolan-2-yl)pyridine (7): A solution of6-bromopicolinaldehyde (6, 10.0 g, 53.8 mmol), ethylene glycol (16.7 g,26.9 mmol), para-toluenesulfonic acid (100 mg) in dry toluene (250 mL)was refluxed for 3 h while using a Dean-Stark apparatus. The solutionwas concentrated in vacuo and the remaining dark yellow oil was purifiedby silica gel chromatography eluting with 0-40% EtOAc in hexanes toafford the title compound (10.4 g, 84%) as a clear, colorless oil: ¹HNMR (500 MHz, CDCl₃) δ 7.59 (t, J=7.5 Hz, 1H), 7.53-7.44 (m, 2H), 7.25(d, J=8.0 Hz, 1H), 5.81 (s, 1H), 4.25-4.03 (m, 4H).

Preparation of1-[6-(1,3-Dioxolan-2-yl)pyridin-2-yl]-4-isopropylpiperazine (8): Amixture of 2-bromo-6-(1,3-dioxolan-2-yl)pyridine (7, 0.23 g, 1.0 mmol)and 1-isopropylpiperazine (0.38 g, 3.0 mmol) in a sealed tube was heatedat 120° C. for 18 h. The mixture was cooled to room temperature, dilutedwith ethyl acetate (50 mL), washed with water (10 mL), brine (10 mL),dried (Na₂SO₄), filtered, and concentrated. The residue was purified bysilica gel chromatography eluting with 0-10% methanol in dichloromethaneto afford the title compound (0.15 g, 55%) as a yellow oil: ¹H NMR (500MHz, CDCl₃) δ 7.51-7.48 (m, 1H), 6.83-6.82 (m, 1H), 6.62-6.60 (m, 1H),5.72 (s, 1H), 4.18-4.14 (m, 2H), 4.08-4.04 (m, 2H), 3.58 (br s, 4H),2.71 (br s, 1H), 2.62 (br s, 2H), 1.08 (d, J=5.5 Hz, 6H); ESI MS m/z 278[M+H]⁺.

Preparation of 6-(4-Isopropylpiperazin-1-yl)picolinaldehyde (9): Amixture of 1-[6-(1,3-dioxolan-2-yl)pyridin-2-yl]-4-isopropylpiperazine(8, 0.31 g, 1.1 mmol), formic acid (2.50 g, 50 mmol) and water (0.5 mL)was heated at 60° C. for 5 h. After this time, the mixture wasconcentrated and the residue was purified by silica gel chromatographyeluting with 0-10% methanol in dichloromethane to afford the titlecompound (0.21 g, 82%) as a yellow oil; ESI MS m/z 234 [M+H]⁺.

Preparation of2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxy-7-phenylquinazolin-4(3H)-one(Example 25): A solution of3-amino-5-methoxy-[1,1′-biphenyl]-4-carboxamide (5, 0.048 g, 0.2 mmol)and 6-(4-isopropylpiperazin-1-yl)picolinaldehyde (9, 0.046 g, 0.2 mmol)in DMA (2 mL) was treated with p-TsOH (0.084 g, 0.44 mmol) and NaHSO₃(0.042 g, 0.5 mmol) and then heated at 120° C. for 3 days. After thistime, the reaction mixture was cooled to room temperature, diluted withsaturated NaHCO₃, extracted with CH₂Cl₂ (2×15 mL), dried (MgSO₄),filtered, and concentrated. The residue was purified by silica gelchromatography eluting with 0-20% methanol in dichloromethane to affordthe title compound (0.017 g, 19%) as a white solid: ¹H NMR (500 MHz,CDCl₃) δ 11.16 (br s, 1H), 7.88-7.86 (m, 2H), 7.78-7.73 (m, 2H),7.55-7.53 (m, 3H), 7.49-7.46 (m, 1H), 7.28-7.27 (m, 1H), 7.09-7.07 (m,1H), 4.08 (s, 3H), 3.64 (br s, 4H), 2.60 (m, 1H), 2.46 (s, 4H), 0.98 (brs, 6H); ESI MS m/z 456 [M+H]⁺.

Example 26 Preparation of8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5-methoxyquinazolin-4(3H)-one

Preparation of 2-(3-Bromophenyl)-1,3-dioxane (2): A solution of3-bromobenzaldehyde (1, 18.5 g, 100 mmol), propane-1,3-diol (9.1 g, 120mmol) and p-toluenesufonic acid monohydrate (0.1 g, 0.5 mmol) in toluene(200 mL) was refluxed with azeotropical removal of water using aDean-Stark water separator for 15.5 h. The reaction mixture was cooledto room temperature, washed with 5% potassium carbonate solution (100mL) and water (2×100 mL), dried over sodium sulfate and filtered. Thefiltrate was concentrated to afford the title compound (24.8 g, 100%):¹H NMR (500 MHz, CDCl₃) δ 7.66 (t, J=1.8 Hz, 1H), 7.46 (ddd, J=7.9, 2.0,1.1 Hz, 1H), 7.40 (br d, J=7.9 Hz, 1H), 7.23 (t, J=7.9 Hz, 1H), 5.46 (s,1H), 4.28-4.25 (m, 2H), 4.00-3.95 (m, 2H), 2.26-2.17 (m, 1H), 1.47-1.43(m, 1H); ESI MS m/z 243, 245 [M+H]⁺.

Preparation of 3-(4-Isopropylpiperazin-1-yl)benzaldehyde (3): A mixtureof 2-(3-bromophenyl)-1,3-dioxane (2, 1.58 g, 6.5 mmol),1-isopropylpiperazine (1.0 g, 7.8 mmol), rac-BINAP (60 mg, 0.096 mmol),and tert-BuONa (1.06 g, 11.1 mmol) in toluene (15 mL) was degassed undervacuum and flushed with nitrogen. Pd₂(dba)₃ (30 mg, 0.033. mmol) wasadded. The reaction mixture was degassed again and flushed with nitrogenand was heated at 100° C. under nitrogen for 16 h. The reaction mixturewas poured into cold 1N HCl (30 mL) and stirred for 2 h. It was adjustedto pH 8 with 6N NaOH and extracted with EtOAc (3×50 mL). The combinedorganic layers were dried (MgSO₄), filtered, concentrated, and purifiedby silica gel chromatography eluting with 0-100% EtOAc (containing 5%v/v Et₃N) in hexanes to afford the title compound as a viscous yellowoil (1.32 g, 87%): ¹H NMR (500 MHz, CDCl₃) δ 9.96 (s, 1H), 7.44-7.37 (m,2H), 7.34-7.30 (m, 1H), 7.21-7.16 (m, 1H), 3.28 (t, J=5.0 Hz, 4H), 2.73(sept, J=6.5 Hz, 1H), 2.69 (t, J=5.0 Hz, 4H), 1.10 (d, J=6.5 Hz, 6H);ESI MS m/z 233 [M+H]⁺.

Preparation of 4-Methoxyphenyl Methyl Carbonate (5): To a stirredsolution of 4-methoxyphenol (4, 12.4 g, 100 mmol) and pyridine (8.5 mL,105 mmol) in CH₂Cl₂ (75 mL) at 0° C. was added ClCO₂Me (8.1 mL, 105mmol) dropwise. The reaction mixture was allowed to warm to roomtemperature and stirred at room temperature for 1 h. The mixture waswashed with 1N HCl, saturated NaHCO₃ solution and water, dried (Na₂SO₄),filtered, and concentrated. The residue was subjected to vacuumdistillation (bp 84-92° C./ca. 1 mmHg) to afford the title compound(16.5 g, 91%) as a colorless oil: ¹H NMR (500 MHz, CDCl₃) δ 7.09 (J=9.1Hz, 2H), 6.89 (d, J=9.1 Hz, 2H), 3.89 (s, 3H), 3.80 (s, 3H).

Preparation of 3-Formyl-4-methoxyphenyl Methyl Carbonate (6): To astirred solution of 4-methoxyphenyl methyl carbonate (5, 16.5 g. 91mmol) in CH₂Cl₂ (205 mL) at 0° C. was added dropwise TiCl₄(23.5 mL, 210mmol) in CH₂Cl₂ (20 mL) over 30 min. Then MeOCHCl₂ (9.3 mL, 104 mmol) inCH₂Cl₂ (20 mL) was added dropwise over 30 min. The reaction mixture wasallowed to warm to room temperature and stirred for 30 min. The reactionmaterial was poured into a mixture of ice (210 g) and concentrated HCl(8.5 mL). EtOAc (200 mL) was added and the mixture was stirredvigorously for 30 min. The aqueous layer was extracted with EtOAc (2×100mL). The combined organic layers were dried (Na₂SO₄), filtered, andconcentrated. The residue was recrystallized from EtOAc/hexanes toafford the title compound (16.6 g, 87%) as a light yellow solid: ¹H NMR(500 MHz, CDCl₃) δ 10.43 (s, 1H), 7.63 (d, J=3.1 Hz, 1H), 7.37 (dd,J=9.0, 3.1 Hz, 1H), 7.01 (d, J=9.0 Hz, 1H), 3.94 (s, 3H), 3.90 (s, 3H);ESI MS m/z 228 [M+H₂O]⁺.

Preparation of 5-(Benzyloxy)-2-methoxybenzaldehyde (7): To a stirredsolution of sodium methoxide (257 mg. 4.8 mmol) in MeOH (20 mL) at roomtemperature was added 3-formyl-4-methoxyphenyl methyl carbonate (6, 1.0g, 4.8 mmol). The resulting orange solution was stirred for 45 min andconcentrated to dryness. The residue was dissolved in DMF (2 mL) andtreated with K₂CO₃ (1.3 g, 9.4 mmol) and BnBr (0.57 mL, 4.8 mL). Thereaction mixture was stirred for 15 h and it was poured into water (20mL). The precipitate was collected, washed with water and dried in vacuoto afford the title compound (1.13 g, 98%) as a light yellow solid: ¹HNMR (500 MHz, CDCl₃) δ 10.44 (s, 1H), 7.49-7.41 (m, 3H), 7.40-7.36 (m,2H), 7.35-7.30 (m, 1H), 7.21 (dd, J=9.1, 3.2 Hz, 1H), 6.95 (d, J=9.1 Hz,1H), 5.05 (s, 2H), 3.90 (s, 3H); ESI MS m/z 243 [M+H]⁺.

Preparation of 5-(Benzyloxy)-2-methoxybenzoic Acid (8): To a stirredsuspension of 5-(benzyloxy)-2-methoxybenzaldehyde (7, 1.12 g. 4.6 mmol)in MeOH (8 mL) at room temperature was added a solution of KOH (1.04 g,18.5 mmol) in water (1.4 mL). The mixture was heated to 65° C. and H₂O₂(35 wt %; 4.0 mL, 46 mmol) was added dropwise over 105 min. The reactionmixture was stirred at 65° C. for 1 h. Then it was cooled to roomtemperature and acidified to pH 2 with 6N H₂SO₄. The precipitate wascollected, washed with water and dried in vacuo to afford the titlecompound (1.16 g, 97%) as a white solid: ¹H NMR (500 MHz, CDCl₃) δ 10.94(br s, 1H), 7.80 (d, J=3.3 Hz, 1H), 7.44-7.37 (m, 4H), 7.35-7.32 (m,1H), 7.19 (dd, J=9.0, 3.3 Hz, 1H), 7.00 (d, J=9.0 Hz, 1H), 5.08 (s, 2H),4.04 (s, 3H); ESI MS m/z 257 [M−H]⁻.

Preparation of 5-(Benzyloxy)-2-methoxybenzamide (9): To a stirredsolution of 5-(benzyloxy)-2-methoxybenzoic acid (8, 1.16 g. 4.5 mmol) inCH₂Cl₂ (25 mL) at room temperature was added oxalyl chloride (1.5 mL,17.9 mmol). The reaction mixture was stirred at room temperature for 2h. It was concentrated to dryness to provide the crude acid chloride asa brown-yellow waxy solid. The crude acid chloride was dissolved in THF(10 mL) and was added dropwise to a stirred mixture of concentratedNH₄OH (20 mL) and THF (10 mL) at 0° C. The reaction mixture was warmedto room temperature and stirred for 1 h and was adjusted to pH 8 with 2N HCl. The precipitate was collected, washed with water, and dried invacuo to afford the title compound (0.73 g, 63%) as a white solid: ¹HNMR (500 MHz, CDCl₃) δ 7.88 (d, J=3.3 Hz, 1H), 7.82 (br s, 1H),7.47-7.43 (m, 2H), 7.40-7.37 (m, 2H), 7.33-7.30 (m, 1H), 7.10 (dd,J=9.0, 3.3 Hz, 1H), 6.94 (d, J=9.0 Hz, 1H), 5.75 (br s, 1H), 5.08 (s,2H), 3.94 (s, 3H); ESI MS m/z 258 [M+H]⁺.

Preparation of 3-(Benzyloxy)-6-methoxy-2-nitrobenzamide (10): To astirred mixture of 5-(benzyloxy)-2-methoxybenzamide (9, 0.365 g. 1.4mmol) and Ac₂O (1.2 mL) at 0° C. was added 70% HNO₃ (0.14 mL, 2.1 mmol).The reaction mixture was warmed to room temperature and stirred for 1 h.Then 4.5 mL of 70% HNO₃ precooled at −30° C. was added. The mixture wasstirred at 0° C. for 1 h and was poured onto ice. The precipitate wascollected, washed with water and dried in vacuo. Purification by silicagel chromatography eluting with 0-2.5% MeOH in CH₂Cl₂ afforded the titlecompound along with its regioisomer (0.34 g, 79%) as a yellow solid: ¹HNMR (500 MHz, CDCl₃) δ 7.73 (br s, 1H), 7.41-7.30 (m, 5H), 7.11 (d,J=9.2 Hz, 1H), 7.00 (d, J=9.2 Hz, 1H), 6.84 (br s, 1H), 5.15 (s, 2H),3.91 (s, 3H); ESI MS m/z 303 [M+H]⁺.

Preparation of 2-Amino-3-(benzyloxy)-6-methoxybenzamide (11): A mixtureof 3-(benzyloxy)-6-methoxy-2-nitrobenzamide (10, 0.34 g. 1.1 mmol), ironpowder (0.44 g, 7.7 mmol) and concentrated hydrochloric acid (80 mL) inEtOH (4 mL)/water (0.4 mL) was heated at 90° C. for 75 min. The reactionmixture was cooled to room temperature, filtered through Celite andconcentrated. The residue was dissolved in EtOAc washed with saturatedNaHCO₃ solution, dried over Na₂SO₄, filtered, and concentrated.Purification by silica gel chromatography eluting with 0-10% MeOH inCH₂Cl₂ afforded the title compound along with its regioisomer (0.267 g,87%) as a tan solid: ¹H NMR (500 MHz, CDCl₃) δ 7.83 (br s, 1H),7.47-7.32 (m, 5H), 6.77 (d, J=8.8 Hz, 1H), 6.60 (br s, 2H), 6.06 (d,J=8.8 Hz, 1H), 5.51 (br s, 1H), 5.04 (s, 2H), 3.85 (s, 3H).

Preparation of8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5-methoxyquinazolin-4(3H)-one(Example 26): A solution of 2-amino-3-(benzyloxy)-6-methoxybenzamide(11, 0.267 g, 0.98 mmol) and 5-(benzyloxy)-2-methoxybenzaldehyde (3,0.152 g, 0.65 mmol) in N,N-dimethylacetamide (10 mL) was treated withp-TsOH (0.30 g, 1.57 mmol) and NaHSO₃ (0.204 g, 1.96 mmol) and thenheated at 110° C. for 15 h. The reaction mixture was cooled to roomtemperature, and concentrated under reduced pressure. The residue waspurified by silica gel chromatography eluting with 0-100% 95:4.5:0.5CH₂Cl₂/MeOH/concentrated NH₄OH in CH₂Cl₂ to afford the title compound(71 mg, 22%) as a light yellow solid: ¹H NMR (500 MHz, DMSO-d₆) δ 12.22(s, 1H), 7.79 (s, 1H), 7.68 (d, J=7.6 Hz, 1H), 7.56 (d, J=7.3 Hz, 2H),7.42-7.33 (m, 5H), 7.14 (d, J=7.6 Hz, 1H), 6.92 (d, J=9.0 Hz, 1H), 5.24(s, 2H), 3.83 (s, 3H), 3.34-3.31 (m, 4H), 2.80-2.60 (m, 5H), 1.05 (br s,6H); ESI MS m/z 485 [M+H]⁺.

Example 27 Preparation of4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-isopropylpiperazine-1-carboxamideHydrochloride

Preparation of Benzyl 4-(Isopropylcarbamoyl)piperazine-1-carboxylate(3): To a solution of benzyl piperazine-1-carboxylate (1, 5.0 g, 23mmol) and diisopropylethylamine (5.86 g, 45.4 mmol) in dichloromethane(50 mL) was added 2-isocyanatopropane (4.83 g, 56.8 mmol) and themixture was stirred overnight at room temperature. The mixture wasdiluted with EtOAc (300 mL), washed with water (100 mL), dried (Na₂SO₄),filtered, and concentrated. The residue was purified by silica gelchromatography eluting with to 0-20% CH₂Cl₂/92:7:1CHCl₃/MeOH/concentrated NH₄OH in CH₂Cl₂ to afford the title compound(3.94 g, 57%) as a white solid: ¹H NMR (300 MHz, CDCl₃) δ 7.45-7.25 (m,5H), 5.14 (s, 2H), 4.19 (d, J=6.7 Hz, 1H), 4.05-3.90 (m, 1H), 3.60-3.45(m, 4H), 3.44-3.25 (m, 4H), 1.20-1.10 (m, 6H); Multimode MS m/z 306[M+H]⁺.

Preparation of N-Isopropylpiperazine-1-carboxamide (4): To a solution ofbenzyl 4-(isopropylcarbamoyl)piperazine-1-carboxylate (3, 3.94 g, 12.9mmol) in methanol (100 mL) was added 10% Pd/C (50% wet, 0.400 g) and themixture was stirred under 1 atmosphere of hydrogen for 16 h. After thistime, the mixture was filtered through Celite and the filtrateconcentrated to afford the title compound (2.88 g, >99%): ¹H NMR (300MHz, CD₃OD) δ 4.00-3.80 (m, 1H), 3.45-3.35 (m, 4H), 2.88-2.78 (m, 4H),1.13 (d, J=6.5 Hz, 6H); Multimode MS m/z 172 [M+H]⁺.

Preparation of4-(6-Formylpyridin-2-yl)-N-isopropylpiperazine-1-carboxamide (6): Amixture of 6-bromopicolinaldehyde (5, 1.00 g, 5.38 mmol) andN-isopropylpiperazine-1-carboxamide (4, 1.10 g, 6.45 mmol) inN,N-dimethylacetamide (20 mL) was treated with K₂CO₃ (2.23 g, 16.1 mmol)and heated at 80° C. for 18 h. The mixture was cooled to roomtemperature and concentrated. The residue was purified by silica gelchromatography eluting with hexanes to 100% EtOAc to 4:1 EtOAc/MeOH toafford the title compound (0.35 g, 24%) as an impure yellow solid:Multimode MS m/z 172 [M+H]⁺.

Preparation of4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-isopropylpiperazine-1-carboxamideHydrochloride (Example 27): A solution of4-(6-formylpyridin-2-yl)-N-isopropylpiperazine-1-carboxamide (6, 0.350g, 1.27 mmol) and 2-amino-4,6-dimethoxybenzamide (7, 0.165 g, 0.845mmol) in DMA (5 mL) was treated with p-TsOH (0.273 g, 1.44 mmol) andNaHSO₃ (0.158 g, 1.52 mmol) and then heated at 100° C. for 16 h. Thereaction mixture was cooled to room temperature and concentrated. Theresidue was purified by silica gel chromatography eluting with 0-50%CH₂Cl₂/92:7:1 CHCl₃/MeOH/concentrated NH₄OH in CH₂Cl₂. A portion of thismaterial was further purified by reverse phase HPLC eluting with 10% to90% CH₃CN in H₂O with 0.1% TFA. The product fractions were combined andconcentrated. The residue was dissolved in water, acidified with 5 N HCland concentrated to afford the title compound in hydrochloric acid saltform (0.042 g, 9%) as a brown solid: ¹H NMR (300 MHz, CD₃OD) δ 7.88 (dd,J=7.4, 8.7 Hz, 1H), 7.66 (d, I=7.3 Hz, 1H), 7.27 (d, J=7.3 Hz, 1H), 7.03(d, J=2.2 Hz, 1H), 6.82 (d, J=2.2 Hz, 1H), 4.01 (s, 3H), 4.01 (s, 3H),3.85-4.00 (m, 1H), 3.75-3.83 (m, 4H), 3.52-3.62 (m, 4H), 1.17 (d, J=6.6Hz, 6H); ESI MS m/z 453 [M+H]⁺.

Example 28 Preparation of2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride

Preparation of 4-isopropylpiperazin-2-one (2): To a solution ofpiperazin-2-one (1, 0.5 g, 5 mmol) and acetone (0.58 g, 10 mmol) in1,2-dichloroethane (20 mL) was added sodium triacetoxyborohydride (3.18g, 15.0 mmol) and the mixture was heated at 70° C. for 2 h. The mixturewas diluted with EtOAc/saturated Na₂CO₃ (100/50 mL), washed with brine(20 mL), dried (Na₂SO₄), filtered and concentrated to afford the crudetitle compound (0.56 g, 79%).

Preparation of 2-(6-Bromopyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(5): A solution of 6-bromopicolinaldehyde (4, 2.8 g, 15 mmol) and2-amino-4,6-dimethoxybenzamide (3, 2.00 g, 10.2 mmol) in DMA (150 mL)was treated with p-TsOH (4.2 g, 22 mmol) and NaHSO₃ (2.6 g, 25 mmol) andthen heated at 110° C. for 16 h. The reaction mixture was cooled to roomtemperature and concentrated. The residue was purified by silica gelchromatography eluting with 0-2% MeOH in CH₂Cl₂ to afford the titlecompound (1.87 g, 51%) as a yellow solid: ¹H NMR (500 MHz, CDCl₃) δ10.40 (s, 1H), 8.49 (d, J=7.5 Hz, 1H), 7.75 (t, J=7.5 Hz, 1H), 7.65 (d,J=7.5 Hz, 1H), 6.83 (d, J=2.2 Hz, 1H), 6.52 (d, J=2.2 Hz, 1H), 4.00 (s,3H), 3.95 (s, 3H); Multimode MS m/z 362 [M+H]⁺.

Preparation of2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride (Example 28): To a mixture of2-(6-bromopyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (5, 0.320 g,0.886 mmol), 4-isopropylpiperazin-2-one (2, 0.152 g, 1.07 mmol), cesiumcarbonate (0.572 g, 1.76 mmol), and XantPhos (0.024 g, 0.044 mmol) intoluene (100 mL) under nitrogen was added Pd₂(dba)₃(0.040 g, 0.044 mmol)and the mixture heated at 110° C. for 48 h. The mixture was cooled toroom temperature. The reaction mixture was loaded directly on a silicagel column and eluted with 0-2% MeOH in CH₂Cl₂. The material was furtherpurified by reverse phase HPLC eluting with 10% to 90% CH₃CN in H₂O with0.1% TFA. The product fractions were combined and concentrated. Theresidue was dissolved in water, acidified with 5 N HCl, and concentratedto afford the title compound (0.091 g, 22%) as a yellow solid: ¹H NMR(500 MHz, DMSO-d₆) δ 11.38 (s, 1H), 8.25-8.32 (m, 1H), 8.10-8.18 (m,2H), 6.83 (d, J=2.2 Hz, 1H), 6.62 (d, J=2.2 Hz, 1H), 4.89 (d, J=13.5 Hz,1H), 4.35-4.22 (m, 1H), 4.22-4.10 (m, 1H), 4.10-4.00 (m, 1H), 3.95-3.80(m, 1H), 3.60-3.72 (m, 1H), 3.55-3.60 (m, 1H), 3.92 (s, 3H), 3.88 (s,3H), 1.37 (d, J=6.5 Hz, 6H); ESI MS m/z 424 [M+H]⁺.

Example 29 Preparation of2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one

Preparation of2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one(Example 29): To a high pressure vial was added2-(6-bromopyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (5 Scheme 21,100 mg, 0.27 mmol) and N,N-dimethylpiperidin-4-amine (142 mg, 1.10mmol). The tube was sealed and the reaction was heated to 110° C. for 4h. The product was purified by silica gel chromatography eluting with0-5% CH₂Cl₂:CH₃OH:aq. NH₄OH (80/18/2) in CH₂Cl₂ to afford the titlecompound (0.815 g, 31%) as a yellow solid: ¹H NMR (500 MHz, DMSO-d₆) δ10.9 (s, 1H), 7.72 (t, J=8.0 Hz, 1H), 7.64 (d, J=7.0 Hz, 1H), 7.07 (d,J=8.5 Hz, 1H), 6.77 (d, J=2.0 Hz, 1H), 6.57 (d, J=2.0 Hz, 1H), 4.49-4.46(m, 2H), 3.90 (s, 3H), 3.86 (s, 3H), 2.93-2.83 (m, 2H), 2.42-2.30 (m,1H), 2.21 (s, 6H), 1.90-1.82 (m, 2H), 1.48-1.30 (m, 2H); ESI MS m/z 410[M+H]⁺.

Example 30 Preparation of5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one

Preparation of5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one(Example 30): A mixture of2-(6-bromopyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one (5 Scheme 21,0.100 g, 0.278 mmol), 1-methylpiperazin-2-one (7, 0.100 g, 0.667 mmol)and anhydrous lithium hydroxide (0.020 g, 0.83 mmol) in 15-crown-5 (0.9mL) was heated at 100° C. for 18 h. The mixture was cooled to roomtemperature and purified by reverse phase HPLC eluting with 10% to 90%CH₃CN in H₂O to afford the title compound (0.02 g, 18%) as an off-whitesolid: ¹H NMR (500 MHz, DMSO-d₆) δ 11.33 (s, 1H), 7.78 (t, J=7.5 Hz,1H), 7.73 (d, J=7.1 Hz, 1H), 7.08 (d, J=8.3 Hz, 1H), 6.78 (d, J=2.2 Hz,1H), 6.58 (d, J=2.2 Hz, 1H), 4.25 (s, 2H), 3.97 (t, J=5.5 Hz, 2H), 3.91(s, 3H), 3.87 (s, 3H), 3.47 (t, J=5.3 Hz, 2H), 2.93 (s, 3H); ESI MS m/z396 [M+H]⁺.

Example 31 Preparation of2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one

Preparation of tert-Butyl4-(3-Chloro-5-formylphenyl)piperazine-1-carboxylate (3): To a mixture of3-bromo-5-chlorobenzaldehyde (1, 0.250 g, 1.14 mmol), tert-butylpiperazine-1-carboxylate (2, 0.255 g, 1.37 mmol), cesium carbonate(0.667 g, 2.05 mmol), and BINAP (0.106 g, 0.171 mmol) in toluene (100mL) was added Pd₂(dba)₃ (0.104 g, 0.114 mmol). The reaction was placedunder N₂ and heated at 110° C. for 16 h. The mixture was cooled to roomtemperature and was loaded directly on silica gel column and eluted with0-50% EtOAc in hexanes to afford the crude title compound (0.3 g, 81%).

Preparation of2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one(Example 31): A solution of tert-butyl4-(3-chloro-5-formylphenyl)piperazine-1-carboxylate (3, 0.300 g, 0.918mmol) and 2-amino-4,6-dimethoxybenzamide (4, 0.120 g, 0.612 mmol) in DMA(4 mL) was treated with p-TsOH (0.424 g, 1.46 mmol) and NaHSO₃ (0.290 g,1.84 mmol) and then heated at 110° C. for 16 h. The reaction mixture wascooled to room temperature, trifluoroacetic acid (5 mL) was added andthe mixture was heated at 70° C. for 1 h. The reaction mixture wascooled to room temperature and concentrated. The residue was purified bysilica gel chromatography eluting with 0-100% 92:7:1CHCl₃/MeOH/concentrated NH₄OH in CH₂Cl₂. This material was furtherpurified by reverse phase HPLC eluting with 10% to 90% CH₃CN in H₂O with0.1% TFA. The product fractions were concentrated and the residue wasdissolved in acetonitrile and water. The mixture was basified withconcentrated NH₄OH and the precipitate filtered, washed with water, anddried under vacuum to afford the title compound (0.072 g, 29%) as anoff-white solid: ¹H NMR (500 MHz, DMSO-d₆) δ 7.60-7.70 (m, 2H),7.08-7.12 (m, 1H), 6.78 (d, J=2.3 Hz, 1H), 6.55 (d, J=2.3 Hz, 1H), 3.89(s, 3H), 3.85 (s, 3H), 3.24 (t, J=5.2 Hz, 4H), 3.28 (t, J=5.2 Hz, 4H);ESI MS m/z 401 [M+H]⁺.

Example 32 Preparation of2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one

To a solution of2-(3-chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one(Example 31, 0.054 g, 0.14 mmol), acetone (0.060 g, 1.4 mmol), sodiumacetate (0.023 g, 0.27 mmol), and acetic acid (0.16 g, 2.7 mmol) in1,2-dichloroethane/methanol (5/5 mL) was added sodiumtriacetoxyborohydride (0.56 g, 2.7 mmol). The mixture was stirred atroom temperature for 16 h and then concentrated. The residue waspurified by silica gel chromatography eluting with 0-100% 92:7:1CHCl₃/MeOH/concentrated NH₄OH in CH₂Cl₂. This material was furtherpurified by reverse phase HPLC eluting with 10% to 90% CH₃CN in H₂O with0.1% TFA. The product fractions were concentrated and the residue wasdissolved in acetonitrile and water. The mixture was basified withconcentrated NH₄OH and concentrated. The residue was purified by reversephase HPLC eluting with 10% to 90% CH₃CN in H₂O without TFA to affordthe title compound (0.011 g, 18%) as an off-white solid: ¹H NMR (500MHz, CD₃OD) δ 7.52 (s, 1H), 7.49 (d, J=1.5 Hz, 1H), 7.18 (t, J=1.5 Hz,1H), 6.82 (d, J=2.5 Hz, 1H), 6.59 (d, J=2.5 Hz, 1H), 3.93 (s, 3H), 3.92(s, 3H), 3.35-3.60 (br s, 4H), 2.85-3.15 (br s, 4H), 1.25-1.40 (br s,1H), 1.23 (d, J=6.5 Hz, 6H); ESI MS m/z 443 [M+H]⁺.

Example 33 Inhibition of Tetra-Acetylated Histone H4 Binding IndividualBET Bromodomains

Proteins were cloned and overexpressed with a N-terminal 6×His tag, thenpurified by nickel affinity followed by size exclusion chromatography.Briefly, E. coli BL21(DE3) cells were transformed with a recombinantexpression vector encoding N-terminally Nickel affinity taggedbromodomains from Brd2, Brd3, Brd4. Cell cultures were incubated at 37°C. with shaking to the appropriate density and induced overnight withIPTG. The supernatant of lysed cells was loaded onto Ni-IDA column forpurification. Eluted protein is pooled, concentrated and furtherpurified by size exclusion chromatography. Fractions representingmonomeric protein were pooled, concentrated, aliquoted, and frozen at−80° C. for use in subsequent experiments.

Binding of tetra-acetylated histone H4 and BET bromodomains wasconfirmed by a Time Resolved Fluorescence Resonance Energy Transfer(TR-FRET) method. N-terminally His-tagged bromodomains (200 nM) andbiotinylated tetra-acetylated histone H4 peptide (25-50 nM, Millipore)were incubated in the presence of Europium Cryptate-labeled streptavidin(Cisbio Cat. #610SAKLB) and XL665-labeled monoclonal anti-His antibody(Cisbio Cat. #61HISXLB) in a white 96 well microtiter plate (Greiner).For inhibition assays, serially diluted test compound was added to thesereactions in a 0.2% final concentration of DMSO. Final bufferconcentrations were 30 mM HEPES pH 7.4, 30 mM NaCl, 0.3 mM CHAPS, 20 mMphosphate pH 7.0, 320 mM KF, 0.08% BSA). After 2 hours incubation atroom temperature, the fluorescence by FRET was measured at 665 and 620nm by a SynergyH4 plate reader (Biotek). Illustrative results with thefirst bromodomain of Brd4 are shown below. The binding inhibitoryactivity was shown by a decrease in 665 nm fluorescence relative to 620nm. IC₅₀ values were determined from a dose response curve.

Compounds with an IC₅₀ value less than 30 μM were deemed to be active.

TABLE 1 Inhibition of Binding of Tetra-acetylated Histone H4 and Brd4bromodomain 1 as Measured by FRET FRET activity Name (IC50 < 30 μM)2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin- Active4(3H)-one [Example 1]2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 2]5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2- Activeyl)quinazolin-4(3H)-one [Example 3]2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 4]2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one [Example 5]5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-Active yl)quinazolin-4(3H)-one [Example 6] Methyl2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin- Active2-yl)piperazin-1-yl)acetate [Example 7]2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 8]2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-Active yl)piperazin-1-yl)propanamide [Example 9]2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-Active yl)piperazin-1-yl)acetic acid [Example 10]3-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-Active yl)piperazin-1-yl)propanoic acid [Example 11]2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-Active yl)-5,7-dimethoxyquinazolin-4(3H)-one bis(trifluoroacetate)[Example 12]2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-Active dimethoxyquinazolin-4(3H)-one acetate [Example 13]2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 14]5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-Active 4(3H)-one Hydrochloride (Example 15)5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-one ActiveHydrochloride (Example 16)2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one Hydrochloride (Example 17)5,7-Dimethoxy-2-(6-(4-(methylsulfonyl)piperazin-1-yl)pyridin-2- NotActive yl)quinazolin-4(3H)-one Hydrochloride (Example 18)5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-Active 4(3H)-one dihydrochloride (Example 19)2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 20)7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 21)7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 22)7-(4-Isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-Active yl)-5-methoxyquinazolin-4(3H)-one Hydrochloride (Example 23)2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxy-7- Activephenylquinazolin-4(3H)-one (Example 25)8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5- Activemethoxyquinazolin-4(3H)-one (Example 26)4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-Active isopropylpiperazine-1-carboxamide Hydrochloride (Example 27)2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7- Not Activedimethoxyquinazolin-4(3H)-one Hydrochloride (Example 28)2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 29)5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2- Not Activeyl)quinazolin-4(3H)-one (Example 30)2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin- Active4(3H)-one (Example 31)2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 32)

Example 34 Inhibition of c-Myc Expression in Cancer Cell Lines

MV4-11 cells (2.5×10⁴ cells) were plated in 96 well U-bottom plates withtest compound or DMSO (0.1%), and incubated for 3 hours at 37° C. Cellswere then harvested by centrifugation, lysed, and mRNA was isolatedusing the mRNA catcher plus kit (Invitrogen). Reverse transcription ofthe mRNA and duplex amplification of the c-myc and cyclophilin cDNAs wasperformed using the RNA Ultrasense kit (Invitrogen) and a ViiA7real-time PCR machine (Applied Biosystems). IC₅₀ values were determinedfrom a dose response curve.

Compounds with an IC₅₀ value less than 30 μM were deemed to be active.

TABLE 2 Inhibition of c-myc Activity in Human AML MV4-11 cells c-mycactivity Name (IC50 < 30 μM)2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-Active one [Example 1]2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 2]5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2- Activeyl)quinazolin-4(3H)-one [Example 3]2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 4]2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one [Example 5]5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-Not Active yl)quinazolin-4(3H)-one [Example 6]2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 8]2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2- NotActive yl)piperazin-1-yl)propanamide [Example 9]2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-Active yl)-5,7-dimethoxyquinazolin-4(3H)-one bis(trifluoroacetate)[Example 12]2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-Active dimethoxyquinazolin-4(3H)-one acetate [Example 13]2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 14]5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-Active 4(3H)-one Hydrochloride (Example 15)5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-one NotActive Hydrochloride (Example 16)2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one Hydrochloride (Example 17)5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-Not Active 4(3H)-one dihydrochloride (Example 19)2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7- NotActive dimethoxyquinazolin-4(3H)-one (Example 20)7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 21)7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 22)7-(4-Isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-Active yl)-5-methoxyquinazolin-4(3H)-one Hydrochloride (Example 23)8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5- Not Activemethoxyquinazolin-4(3H)-one (Example 26)4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-Active isopropylpiperazine-1-carboxamide Hydrochloride (Example 27)2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 29)5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2- Not Activeyl)quinazolin-4(3H)-one (Example 30)2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin- Active4(3H)-one (Example 31)2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 32)

Example 35 Inhibition of Cell Proliferation in Cancer Cell Lines

MV4-11 cells: 96-well plates were seeded with 5×10⁴ cells per well ofexponentially growing human AML MV-4-11 (CRL-9591) cells and immediatelytreated with two-fold dilutions of test compounds, ranging from 30 μM to0.2 μM. Triplicate wells were used for each concentration, as well as amedia only and three DMSO control wells. The cells and compounds wereincubated at 37° C., 5% CO₂ for 72 hours before adding 20 μL of theCellTiter Aqueous One Solution (Promega) to each well and incubating at37° C., 5% CO₂ for an additional 3-4 hours. The absorbance was taken at490 nm in a spectrophotometer and the percentage of proliferationrelative to DMSO-treated cells was calculated after correction from theblank well. IC₅₀ were calculated using the GraphPad Prism software.

Compounds with an IC₅₀ value less than 30 μM are deemed to be active.

TABLE 3 Inhibition of Cell Proliferation in Human AML MV-4-11 cells cellproliferation activity Name (IC50 < 30 μM)2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin- NotActive 4(3H)-one [Example 1]2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 2]5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2- Activeyl)quinazolin-4(3H)-one [Example 3]2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 4]2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one [Example 5]5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-Not Active 2-yl)quinazolin-4(3H)-one [Example 6]2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 8]2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-Active yl)piperazin-1-yl)propanamide [Example 9]2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-Active yl)-5,7-dimethoxyquinazolin-4(3H)-one bis(trifluoroacetate)[Example 12]2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-Not Active dimethoxyquinazolin-4(3H)-one acetate [Example 13]2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 14]5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-Active 4(3H)-one Hydrochloride (Example 15)5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-one NotActive Hydrochloride (Example 16)2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one Hydrochloride (Example 17)5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-Not Active 4(3H)-one dihydrochloride (Example 19)2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7- NotActive dimethoxyquinazolin-4(3H)-one (Example 20)7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 21)7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 22)7-(4-isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-Active yl)-5-methoxyquinazolin-4(3H)-one Hydrochloride (Example 23)8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5- Not Activemethoxyquinazolin-4(3H)-one (Example 26)4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-Active isopropylpiperazine-1-carboxamide Hydrochloride (Example 27)2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 29)5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2- Not Activeyl)quinazolin-4(3H)-one (Example 30)2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin- Active4(3H)-one (Example 31)2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 32)

Example 36 Inhibition of hIL-6 mRNA Transcription

In this example, hIL-6 mRNA in tissue culture cells was quantitated tomeasure the transcriptional inhibition of hIL-6 when treated with acompound of the present disclosure.

A human leukemic monocyte lymphoma cell line (U937) was plated (3.2×10⁴cells per well) in a 96-well plate in 100 IL RPMI-1640 containing 10%FBS and penicillin/streptomycin, and differentiated into macrophages for3 days in 60 ng/mL PMA (phorbol-13-myristate-12-acetate) at 37° C. in 5%CO₂ prior to the addition of the compound of interest. The cells werepretreated for 1 h with the test compound prior to stimulation with 1ug/mL lipopolysaccharide from Escherichia coli. The cells were incubatedat 37-C for 3 h before the cells were harvested. At time of harvest, thespent media was removed from the cells and the cells were rinsed in 200μL PBS. Cell lysis solution (70 μL) was added the cells in each well andincubated for 5-10 min at room temperature, to allow for complete celllysis and detachment. mRNA was then prepared using the “mRNA CatcherPLUS plate” (Invitrogen), according to the protocol supplied. After thelast wash, as much wash buffer as possible was aspirated withoutallowing the wells to dry. Elution buffer (E3, 70 μL) was then added toeach well. mRNA was then eluted by incubating the mRNA Catcher PLUSplate with Elution Buffer for 5 min at 68° C. and then immediatelyplacing the plate on ice.

The eluted mRNA isolated was then used in a one-step quantitativereal-time PCR reaction, using components of the Ultra Sense Kit togetherwith Applied Biosystems primer-probe mixes. Real-time PCR data wasanalyzed, normalizing the Ct values for hIL-6 to an internal control,prior to determining the fold induction of each unknown sample, relativeto the control.

Compounds with an IC₅₀ value less than 30 μM are deemed to be active.

TABLE 4 Inhibition of hIL-6 mRNA Transcription IL-6 activity Name (IC50< 30 μM)2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-Active one [Example 1]2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 2]5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2- Activeyl)quinazolin-4(3H)-one [Example 3]2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 4]2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one [Example 5]5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-Active yl)quinazolin-4(3H)-one [Example 6] Methyl2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin- Active2-yl)piperazin-1-yl)acetate [Example 7]2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 8]2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2- NotActive yl)piperazin-1-yl)propanamide [Example 9]2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2- NotActive yl)piperazin-1-yl)acetic acid [Example 10]3-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2- NotActive yl)piperazin-1-yl)propanoic acid [Example 11]2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-Not Active yl)-5,7-dimethoxyquinazolin-4(3H)-one bis(trifluoroacetate)[Example 12]2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-Active dimethoxyquinazolin-4(3H)-one acetate [Example 13]2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7- Not Activedimethoxyquinazolin-4(3H)-one [Example 14]5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-Active 4(3H)-one Hydrochloride (Example 15)5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-one ActiveHydrochloride (Example 16)2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one Hydrochloride (Example 17)5,7-Dimethoxy-2-(6-(4-(methylsulfonyl)piperazin-1-yl)pyridin-2- NotActive yl)quinazolin-4(3H)-one Hydrochloride (Example 18)5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-Not Active 4(3H)-one dihydrochloride (Example 19)2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 20)7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 21)7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5- Activemethoxyquinazolin-4(3H)-one Hydrochloride (Example 22)7-(4-Isopropylpiperazin-1-yl)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-Not Active yl)-5-methoxyquinazolin-4(3H)-one Hydrochloride (Example 23)2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxy-7- Not Activephenylquinazolin-4(3H)-one (Example 25)8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5- Not Activemethoxyquinazolin-4(3H)-one (Example 26)4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-Active isopropylpiperazine-1-carboxamide Hydrochloride (Example 27)2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 29)5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2- Activeyl)quinazolin-4(3H)-one (Example 30)2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin- Active4(3H)-one (Example 31)2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 32)

Example 37 Inhibition of hVCAM mRNA Transcription

In this example, hVCAM mRNA in tissue culture cells is quantitated tomeasure the transcriptional inhibition of hVCAM when treated with acompound of the present disclosure.

Human umbilical vein endothelial cells (HUVECs) are plated in a 96-wellplate (4.0×10³ cells/well) in 100 μL EGM media and incubated for 24 hprior to the addition of the compound of interest. The cells arepretreated for 1 h with the test compound prior to stimulation withtumor necrosis factor-α. The cells are incubated for an additional 24 hbefore the cells are harvested. At time of harvest, the spent media isremoved from the HUVECs and rinsed in 200 μL PBS. Cell lysis solution(70 μL) is then added the cells in each well and incubated for ¹⁸ 5-10min at room temperature, to allow for complete cell lysis anddetachment, mRNA is then prepared using the “mRNA Catcher PLUS plate”(Invitrogen), according to the protocol supplied. After the last wash,as much wash buffer as possible is aspirated without allowing the wellsto dry. Elution buffer (E3, 70 μL) is then added to each well. mRNA isthen eluted by incubating the mRNA Catcher PLUS plate with elutionbuffer for 5 min at 68° C. and then immediately placing the plate onice.

The eluted mRNA so isolated is then used in a one-step quantitativereal-time PCR reaction, using components of the Ultra Sense Kit togetherwith Applied Biosystems primer-probe mixes. Real-time PCR data wasanalyzed, normalizing the Ct values for hVCAM to an internal control,prior to determining the fold induction of each unknown sample, relativeto the control.

Compounds with an IC₅₀ value less than 30 μM are deemed to be active.

Example 38 Inhibition of hMCP-1 mRNA Transcription

In this example, hMCP-1 mRNA in human peripheral blood mononuclear cellswas quantitated to measure the transcriptional inhibition of hMCP-1 whentreated with a compound of the present disclosure.

Human Peripheral Blood Mononuclear Cells were plated (1.0×10⁵ cells perwell) in a 96-well plate in 45 μL RPMI-1640 containing 10% FBS andpenicillin/streptomycin. The cells were treated with the test compound(45 μL at 2× concentration), and then the cells were incubated at 37° C.for 3 h before the cells were harvested. At time of harvest, cells weretransferred to V-bottom plates and centrifuged at 800 rpm for 5 minutes.Spent media was removed and cell lysis solution (70 μL) was added thecells in each well and incubated for 5-10 min at room temperature, toallow for complete cell lysis and detachment. mRNA was then preparedusing the “mRNA Catcher PLUS plate” (Invitrogen), according to theprotocol supplied. After the last wash, as much wash buffer as possiblewas aspirated without allowing the wells to dry. Elution buffer (E3, 70μL) was then added to each well. mRNA was then eluted by incubating themRNA Catcher PLUS plate with Elution Buffer for 5 min at 68° C. and thenimmediately placing the plate on ice.

The eluted mRNA isolated was then used in a one-step quantitativereal-time PCR reaction, using components of the Ultra Sense Kit togetherwith Applied Biosystems primer-probe mixes. Real-time PCR data wasanalyzed, normalizing the Ct values for hMCP-1 to an internal control,prior to determining the fold induction of each unknown sample, relativeto the control.

Compounds with an IC₅₀ value less than 30 μM are deemed to be active.

TABLE 5 Inhibition of hMCP-1 mRNA Transcription MCP-1 activity Name(IC50 < 30 μM) 2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one Hydrochloride (Example 17)

Example 39 Up-Regulation of hApoA-1 mRNA Transcription

In this example, ApoA-I mRNA in tissue culture cells was quantitated tomeasure the transcriptional up-regulation of ApoA-I when treated with acompound of the present disclosure.

Huh7 cells (2.5×10⁵ per well) were plated in a 96-well plate using 100μL DMEM per well, (Gibco DMEM supplemented with penicillin/streptomycinand 10% FBS), 24 h before the addition of the compound of interest.After 48 hrs treatment, the spent media was removed from the Huh-7 cellsand placed on ice (for immediate use) or at −80° C. (for future use)with the “LDH cytotoxicity assay Kit II” from Abcam. The cells remainingin the plate were rinsed with 100 μL PBS.

Then 85 μL of cell lysis solution was added to each well and incubatedfor 5-10 minutes at room temperature, to allow for complete cell lysisand detachment. mRNA was then prepared using the “mRNA Catcher PLUSplate” from Life Technologies, according to the protocol supplied. Afterthe last wash, as much wash buffer as possible was aspirated withoutallowing the wells to dry. Elution Buffer (E3, 80 μL) was then added toeach well. mRNA was then eluted by incubating the mRNA Catcher PLUSplate with Elution Buffer for 5 minutes at 68° C., and then 1 minute at4° C. Catcher plates with mRNA eluted were kept on ice for use or storedat −80° C.

The eluted mRNA isolated was then used in a one-step real-time PCRreaction, using components of the Ultra Sense Kit together with LifeTechnologies primer-probe mixes. Real-time PCR data was analyzed, usingthe Ct values, to determine the fold induction of each unknown sample,relative to the control (that is, relative to the control for eachindependent DMSO concentration).

Compounds with an EC₁₇₀ value less than 30 μM are deemed to be active.

TABLE 6 Up-regulation of hApoA-1 mRNA Transcription. ApoA-1 activityName (EC₁₇₀ < 30 μM)2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-Active one [Example 1]2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 2]5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2- Activeyl)quinazolin-4(3H)-one [Example 3]2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 4]2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one [Example 5]5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-Active yl)quinazolin-4(3H)-one [Example 6]2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 8]2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-Active yl)piperazin-1-yl)propanamide [Example 9]3-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2- NotActive yl)piperazin-1-yl)propanoic acid [Example 11]2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-Active yl)-5,7-dimethoxyquinazolin-4(3H)-one bis(trifluoroacetate)[Example 12]2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-Active dimethoxyquinazolin-4(3H)-one acetate [Example 13]2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one [Example 14]5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-Active 4(3H)-one Hydrochloride (Example 15)2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-Active 4(3H)-one Hydrochloride (Example 17)5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-Active 4(3H)-one dihydrochloride (Example 19)2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7- Not Activedimethoxyquinazolin-4(3H)-one Hydrochloride (Example 28)2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 29)2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin- Active4(3H)-one (Example 31)2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7- Activedimethoxyquinazolin-4(3H)-one (Example 32)

Examples 40 In Vivo Efficacy in Athymic Nude Mouse Strain of an AcuteMyeloid Leukemia Xenograft Model Using MV4-11 Cells

MV4-11 cells (ATCC) were grown under standard cell culture conditionsand (NCr) nu/nu fisol strain of female mice age 6-7 weeks were injected5e⁶ cells/animal in 100 μl PBS+100 μl Matrigel in lower left abdominalflank. Approximately by day 18 after MV4-11 cells injection, mice wererandomized based on tumor volume (L×W×H)/2) of average ^(˜)120 mm³. Micewere dosed orally with compound at 75 mg/kg b.i.d and 120 mg/kg b.i.d inEA006 formulation at 10 mL/kg body weight dose volume. Tumormeasurements were taken with electronic micro caliper and body weightsmeasured on alternate days beginning from dosing period. The averagetumor volumes, percent Tumor Growth Inhibition (TGI) and % change inbody weights were compared relative to Vehicle control animals. Themeans, statistical analysis and the comparison between groups werecalculated using student's t-test in Excel.

TABLE 7 In vivo efficacy in athymic nude mouse strain of an acutemyeloid leukemia xenograft model using MV4-11 cells: Name In vivoactivity 2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one Hydrochloride (Example 17)

Example 41 In Vivo Efficacy in Mouse Endotoxemia Model Assay

Sub lethal doses of Endotoxin (E. Coli bacterial lipopolysaccharide)were administered to animals to produce a generalized inflammatoryresponse which was monitored by increases in secreted cytokines.Compounds were administered to C57/Bl6 mice orally at 75 mg/kg dose toevaluate inhibition in IL-6 and IL-17 cytokines post 4 hour challengewith Lipopolysaccharide (LPS) at 0.5 mg/kg dose intraperitoneally.

TABLE 8 In Vivo Efficacy in Mouse Endotoxemia Model Assay. Name In vivoactivity 2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7- Activedimethoxyquinazolin-4(3H)-one Hydrochloride (Example 17)

Example 42 In Vivo Efficacy in Rat Collagen Induced Arthritis

Rat collagen-induced arthritis is an experimental model of polyarthritisthat has been widely used for preclinical testing of numerousanti-arthritic agents. Following administration of collagen, this modelestablishes a measurable polyarticular inflammation, marked cartilagedestruction in association with pannus formation and mild to moderatebone resorption and periosteal bone proliferation. In this model,collagen was administered to female Lewis strain of rats on Day 1 and 7of study and dosed with compounds from Day 11 to Day 17. Test compoundswere evaluated to assess the potential to inhibit the inflammation(including paw swelling), cartilage destruction and bone resorption inarthritic rats, using a model in which the treatment is administeredafter the disease has been established.

TABLE 9 In Vivo Efficacy in Rat Collagen Induced Arthritis. Name In vivoactivity 2-(6-(4-Isopropylpiperazin-1-yl)pyridin- Active2-yl)-5,7-dimethoxyquinazolin-4(3H)-one Hydrochloride (Example 17)

Example 43 In Vivo Efficacy in Experimental Autoimmune Encephalomyelitis(EAE) Model of MS

Experimental autoimmune encephalomyelitis (EAE) is a T-cell-mediatedautoimmune disease of the CNS which shares many clinical andhistopathological features with human multiple sclerosis (MS). EAE isthe most commonly used animal model of MS. T cells of both Th1 and Th17lineage have been shown to induce EAE. Cytokines IL-23, IL-6 and IL-17,which are either critical for Th1 and Th17 differentiation or producedby these T cells, play a critical and non-redundant role in EAEdevelopment. Therefore, drugs targeting production of these cytokinesare likely to have therapeutic potential in treatment of MS.

This study was conducted to assess the potential anti-inflammatoryeffect of test compounds to inhibit the inflammation and clinical EAEscores of a 28 day preventative mouse model. In this model, EAE wasinduced by MOG₃₅₋₅₅/CFA immunization and pertussis toxin injection infemale C57Bl/6 mice.

TABLE 10 In Vivo Efficacy in Experimental autoimmune encephalomyelitis(EAE) Model of MS Name In vivo activity2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)- Active5,7-dimethoxyquinazolin-4(3H)-one Hydro- chloride (Example 17)

Other embodiments of the present disclosure will be apparent to thoseskilled in the art from consideration of the specification and practiceof the present disclosure disclosed herein. It is intended that thespecification and examples be considered as exemplary only, with a truescope and spirit of the present disclosure being indicated by thefollowing claims.

What is claimed is:
 1. A method of inhibiting BET proteins in a mammalby administering a therapeutically effective amount of a compound ofFormula I:

or a stereoisomer, tautomer, pharmaceutical acceptable salt, or hydratethereof, wherein: W₁ is selected from N and CR₁; W₂ is selected from Nand CR₂; W₃ is selected from N and CR₃; W₄ is selected from N and CR₄;each W may be the same or different from each other; A is selected fromN and CH; R₁ and R₄ are each independently selected from hydrogen,alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryloxy, aryl, amino,hydroxyl, and halogen; R₂ and R₃ are each independently selected fromhydrogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryloxy, aryl,hydroxyl, and halogen; two adjacent substituents selected from R₁, R₂,R₃, and R₄ may be connected in a 5- or 6-membered ring to form abicyclic carbocycle or bicyclic heterocycle; AR1 is a group selectedfrom the following:

and

B is a group selected from the following:

each ring system may be substituted with one or more substituentsindependently selected from R₁₀ and R₁₁: R₅ is selected from hydrogen,alkoxy, alkyl, thioalkyl, aryloxy, aryl, hydroxyl, and halogen; R₆ isselected from hydrogen, alkoxy, alkyl, thioalkyl, aryloxy, aryl, andhalogen; R₇ is selected from hydrogen, alkyl, —SO₂R₁₂, —C(O)NR₁₂R₁₃, and—C(O)R₁₂; R₈ and R₉ are independently selected from hydrogen, aryl,alkenyl, alkyl, —SO₂R₁₂, —C(O)NR₁₂R₁₃, and —C(O)R₁₂; R₁₀ and R₁₁ areindependently selected from hydrogen, halogen, alkyl, alkoxy, aryl, andhydroxyl; R₁₂ and R₁₃ are independently selected from hydrogen, aryl,and alkyl; Y is selected from NH, O, and S; and two adjacentsubstituents selected from R₅, R₆, R₈, R₉, R₁₀, and R₁₁ may be connectedin a 5- or 6-membered ring to form a carbocycle or heterocycle.
 2. Themethod according to claim 1, wherein R₁ and R₄ are independentlyselected from hydrogen, alkyl, alkoxy, halogen, and amino; wherein R₂and R₃ are independently selected from hydrogen, alkyl, alkoxy, andhalogen; wherein at least one of R₁, R₂, R₃, and R₄ is not hydrogen;wherein R₅ is selected from hydrogen, alkyl, alkoxy, and halogen;wherein R₆ is selected from hydrogen and alkoxy optionally substitutedwith a hydroxyl or amino; and wherein Y is N.
 3. The compound accordingto claim 2, wherein AR1 is selected from

wherein B is selected from

wherein R₇ is selected from hydrogen and alkyl; wherein R₈ and R₉ areindependently selected from hydrogen and alkyl; wherein R₁₀ and R₁₁ areindependently selected from hydrogen and halogen; and wherein R₁₂ andR₁₃ are independently selected from hydrogen and alkyl.
 4. The methodaccording to claim 1, wherein the compound is selected from:2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;Methyl2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetate;2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanamide;2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)aceticacid;3-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoicacid;2-(5-(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-onebis(trifluoroacetate);2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneacetate;2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-oneHydrochloride;5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-oneHydrochloride;2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride;5,7-Dimethoxy-2-(6-(4-(methylsulfonyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-oneHydrochloride;5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-onedihydrochloride;2-(3-(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride;7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-oneHydrochloride;2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxy-7-phenylquinazolin-4(3H)-one;8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5-methoxyquinazolin-4(3H)-one;4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-isopropylpiperazine-1-carboxamideHydrochloride;2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride;2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;2-(3-Chloro-5-(piperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;and stereoisomers, tautomers, pharmaceutically acceptable salts, orhydrates thereof.
 5. The method according to claim 1, wherein R₆ isselected from the group represented by Formula II:

wherein: D is selected from O and S; E is selected from O, N, and S; R₁₄and R₁₅ are independently selected from hydrogen, alkyl, and cycloalkyl,wherein if E is O or S, only one of R₁₄ and R₁₅ is present; and n isselected from 1, 2, and
 3. 6. The method according to claim 5 wherein Dis O; wherein n=1; wherein R₁₄ and R₁₅ are independently selected fromhydrogen, and alkyl; and wherein R₆ is selected from hydrogen, methoxy,


7. The method of claim 1, further comprising treating a disease,disorder, or condition characterized by dysregulation of cell cyclecontrol, inflammatory cytokine expression, viral transcription,hematopoietic differentiation, insulin transcription, or adipogenesis.8. The method of claim 7, wherein the disease, disorder, or condition isan autoimmune disease.
 9. The method of claim 8, wherein the autoimmunedisease is selected from Acute Disseminated Encephalomyelitis,Agammaglobulinemia, Allergic Disease, Ankylosing spondylitis,Anti-GBM/Anti-TBM nephritis, Anti-phospholipid syndrome, Autoimmuneaplastic anemia, Autoimmune hepatitis, Autoimmune inner ear disease,Autoimmune myocarditis, Autoimmune pancreatitis, Autoimmune retinopathy,Autoimmune thrombocytopenic purpura, Behcet's Disease, Bullouspemphigoid, Castleman's Disease, Celiac Disease, Churg-Strauss syndrome,Crohn's Disease, Cogan's syndrome, Dry eye syndrome, Essential mixedcryoglobulinemia, Dermatomyositis, Devic's Disease, Encephalitis,Eosinophlic esophagitis, Eosinophilic fasciitis, Erythema nodosum, Giantcell arteritis, Glomerulonephritis, Goodpasture's syndrome,Granulomatosis with Polyangiitis (Wegener's), Graves' Disease,Guillain-Barre syndrome, Hashimoto's thyroiditis, Hemolytic anemia,Henoch-Schonlein purpura, IgA nephropathy, Inclusion body myositis, TypeI diabetes, Interstitial cystitis, Kawasaki's Disease, Leukocytoclasticvasculitis, Lichen planus, Lupus (SLE), Microscopic polyangitis,Multiple sclerosis, Myasthenia gravis, myositis, Optic neuritis,Pemphigus, POEMS syndrome, Polyarteritis nodosa, Primary biliarycirrhosis, Psoriasis, Psoriatic arthritis, Pyoderma gangrenosum,Relapsing polychondritis, Rheumatoid arthritis, Sarcoidosis,Scleroderma, Sjogren's syndrome, Takayasu's arteritis, Transversemyelitis, Ulcerative colitis, Uveitis, and Vitiligo.
 10. The method ofclaim 7, wherein the disease, disorder, or condition is an inflammatorydisease.
 11. The method of claim 10, wherein the inflammatory disease ordisorder is selected from sinusitis, pneumonitis, osteomyelitis,gastritis, enteritis, gingivitis, appendicitis, irritable bowelsyndrome, tissue graft rejection, chronic obstructive pulmonary disease(COPD), septic shock, toxic shock syndrome, SIRS, bacterial sepsis,osteoarthritis, acute gout, acute lung injury, acute renal failure,burns, Herxheimer reaction, and SIRS associated with viral infections.12. The method of claim 7, wherein the disease, disorder, or conditionis a cardiovascular disease.
 13. The method of claim 7, wherein thedisease, disorder, or condition is insulin resistance diabetes.
 14. Themethod of claim 7, wherein the disease, disorder, or condition is aneurological disorder.
 15. The method of claim 7, wherein the disease,disorder, or condition is HIV.
 16. The method of claim 7, wherein thedisease, disorder, or condition is cancer.
 17. The method according toclaim 16, wherein R₁ and R₄ are independently selected from hydrogen,alkyl, alkoxy, halogen, and amino; wherein R₂ and R₃ are independentlyselected from hydrogen, alkyl, alkoxy, and halogen; wherein at least oneof R₁, R₂, R₃, and R₄ is not hydrogen; wherein R₅ is selected fromhydrogen, alkyl, alkoxy, and halogen; wherein R₆ is selected fromhydrogen and alkoxy optionally substituted with a hydroxyl or amino; andwherein Y is N.
 18. The compound according to claim 17, wherein AR1 isselected from

wherein B is selected from

wherein R₇ is selected from hydrogen and alkyl; wherein R₈ and R₉ areindependently selected from hydrogen and alkyl; wherein R₁₀ and R₁₁ areindependently selected from hydrogen and halogen; and wherein R₁₂ andR₁₃ are independently selected from hydrogen and alkyl.
 19. The methodaccording to claim 16, wherein the compound is selected from:2-(3-(4-Isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(3-(4-Isopropylpiperazin-1-yl)-5-methoxyphenyl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-(2-methoxyethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;2-(6-(4-(2-Hydroxyethyl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(6-(4-Isobutylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-(2-(methylsulfonyl)ethyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one;Methyl2-(4-(6-(5,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)acetate;2-(6-(4-(1-Hydroxypropan-2-yl)piperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2yl)-pyridin-2-yl)piperazin-1-yl)propanamide;2-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)aceticacid;3-(4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)piperazin-1-yl)propanoicacid;2-(5(2-(Isopropylamino)ethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-onebis(trifluoroacetate);2-(5-(2-Hydroxyethoxy)-6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-oneacetate;2-(6-(4-Isopropylpiperazin-1-yl)-5-methoxypyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-methylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-oneHydrochloride;5,7-Dimethoxy-2-(6-morpholinopyridin-2-yl)quinazolin-4(3H)-oneHydrochloride;2-(6-(4-Isopropylpiperazin-1-yl)pyridin-2-yl)5,7-dimethoxyquinazolin-4(3H)-oneHydrochloride;5,7-Dimethoxy-2-(6-(4-methylsulfonyl)piperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-one Hydrochloride;5,7-Dimethoxy-2-(6-(4-propionylpiperazin-1-yl)pyridin-2-yl)quinazolin-4(3H)-onedihydrochloride,2-(3(4-Isopropylpiperazin-1-yl)-5-(trifluoromethoxy)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;7-Fluoro-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinazolin-4(3H)-one Hydrochloride;7-(Benzyloxy)-2-(6-(4-isopropylpiperazin-1-yl)pyridin-2-yl)-5-methoxyquinzolin-4(3H)-one Hydrochloride;2-(6-(4-Isopropylpiperazin-1-yl)pyrdin-2-yl)-5-methoxy-7-phenyiquinazolin-4(3H)-one; 8-(Benzyloxy)-2-(3-(4-isopropylpiperazin-1-yl)phenyl)-5-methoxyquinazolin -4(3)-one;4-(6-(5,7-Dimethoxy-4-oxo-3,4-dihydroquinazolin-2-yl)pyridin-2-yl)-N-isopropylpiperazine-1-carboxamide Hydrochloride;2-(6-(4-Isopropyl-2-oxopiperazin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one Hydrochloride;2-(6-(4-(Dimethylamino)piperidin-1-yl)pyridin-2-yl)-5,7-dimethoxyquinazolin-4(3H)-one;5,7-Dimethoxy-2-(6-(4-methyl-3-oxopiperazin-1-yl)pyridin-2yl)quinazolin-4(3H)-one;2-(3-Chloro-5-(piperazin-1yl)-phenyl)-5,7-dimethoxyquinazolin-4(3H)-one;2-(3-Chloro-5-(4-isopropylpiperazin-1-yl)phenyl)-5,7-dimethoxyquinazolin-4(3H)-one; and stereoisomers, tautomers, pharmaceutically acceptable salts,or hydrates thereof.
 20. The method according to claim 16, wherein R₆ isselected from the group represented by Formula II:

wherein: D is selected from O and S; E is selected from O, N, and S; R₁₄and R₁₅ are independently selected from hydrogen, alkyl, and cycloalkyl,wherein if E is O or S, only one of R₁₄ and R₁₅ is present; and n isselected from 1, 2, and
 3. 21. The method according to claim 20 whereinD is O; wherein n =1; wherein R₁₄ and R₁₅ are independently selectedfrom hydrogen, and alkyl; and wherein R₆ is selected from hydrogen,methoxy,


22. The method of claim 16, wherein the cancer is a midline carcinoma.23. The method of claim 16, wherein the cancer exhibits overexpression,translocation, amplification, or rearrangement of a myc familyoncoproteins.
 24. The method of claim 23, wherein the cancer ischaracterized by overexpression of c-myc.
 25. The method of claim 23,wherein the cancer is characterized by is characterized byoverexpression n-myc.
 26. The method of claim 16, wherein the cancer ischaracterized by recruitment of pTEFb to regulate oncogenes.
 27. Themethod of claim 16, wherein the cancer is characterized by upregulationof at least one of CDK6, Bcl2, TYRO3, MYB and hTERT.
 28. The method ofclaim 16, wherein the cancer is associated with a viral infection. 29.The method of claim 16, wherein the cancer is selected from B-acutelymphocytic leukemia, Burkitt's lymphoma, diffuse large cell lymphoma,multiple myeloma, primary plasma cell leukemia, atypical carcinoid lungcancer, bladder cancer, breast cancer, cervix cancer, colon cancer,gastric cancer, glioblastoma, hepatocellular carcinoma, large cellneuroendocrine carcinoma, medulloblastoma, melanoma, nodular melanoma,neuroblastoma, oesophageal squamous cell carcinoma, osteosarcoma,ovarian cancer, prostate cancer, renal clear cell carcinoma,retinoblastoma, rhabdomyosarcoma, small cell lung carcinoma, NUT midlinecarcinoma, B-cell lymphoma, non-small cell lung cancer, esophagealcancer and head and neck squamous cell carcinoma, chronic lymphocyticleukemia, follicular lymphoma, diffuse large B cell lymphoma withgerminal center phenotype, Burkitt's lymphoma, Hodgkin's lymphoma,follicular lymphomas, activated anaplastic large cell lymphoma, primaryneuroectodermal tumor, pancreatic cancer, adenoid cystic carcinoma,T-cell prolymphocytic leukemia, malignant glioma, thyroid cancer,Barret's adenocarcinoma, hepatoma, pro-myelocytic leukemia, chroniclymphocytic leukemia, and mantle cell lymphoma.
 30. The method of claim16, wherein the compound of Formula I is administered in combinationwith another anticancer agent.
 31. The method of claim 30, wherein theanticancer agent is selected from ABT-737, Azacitidine (Vidaza), AZD1152(Barasertib), AZD2281 (Olaparib), AZD6244 (Selumetinib), BEZ235,Bleomycin Sulfate, Bortezomib (Velcade), Busulfan (Myleran),Camptothecin, Cisplatin, Cyclophosphamide (Clafen), CYT387, Cytarabine(Ara-C), Dacarbazine, DAPT (GSI-IX), Decitabine, Dexamethasone,Doxorubicin (Adriamycin), Etoposide, Everolimus (RAD001), Flavopiridol(Alvocidib), Ganetespib (STA-9090), Gefitinib (Iressa), Idarubicin,Ifosfamide (Mitoxana), IFNa2a (Roferon A), Melphalan (Alkeran),Methazolastone (temozolomide), Metformin, Mitoxantrone (Novantrone),Paclitaxel, Phenformin, PKC412 (Midostaurin), PLX4032 (Vemurafenib),Pomalidomide (CC-4047), Prednisone (Deltasone), Rapamycin, Revlimid(Lenalidomide), Ruxolitinib (INCB018424), Sorafenib (Nexavar), SU11248(Sunitinib), SU11274, Vinblastine, Vincristine (Oncovin), Vinorelbine(Navelbine), Vorinostat (SAHA), and WP1130 (Degrasyn).